When protoscoleces of Echinococcus multilocularis were cultured in vitro, under axenic conditions in either monophasic or diphasic media, segmentation was suppressed in most organisms, some 70–80% of which developed into unsegmented, monozoic forms with a complete set of sexually mature male and female genitalia. The most striking feature of monozoic worms was the large lateral swelling produced by the cirrus sac the effect being to produce organisms with an unusual asymmetric shape. Worms which did not become monozoic either (a) underwent some somatic growth, developed two sets of genitalia and became ‘pseudosogmented’, i.e. with the inter-proglottid membranes absent or poorly defined, or (b) became vesicular or abnormal. The mechanisms which could be involved in the suppression of somatic growth and the induction of the monozoic condition, are examined in terms of cell lineage. The possible significance of these results in understanding the evolution of the cestodes is discussed.

Malaria causes much physical and economic hardship in endemic countries with billions of people at risk. A vaccine would clearly benefit these countries, reducing the requirement for hospital care and the economic impact of infection. Successful immunization with irradiated sporozoites and the fact that repeated exposure to malaria induces partial immunity to infection and high levels of protection against the clinical manifestations, suggest that a vaccine is feasible. Numerous candidate antigens have been identified but the vaccine, which has been promised to be ‘just round the corner’ for many years, remains elusive. The factors contributing to this frustratingly slow progress are discussed including gaps in the knowledge of host/parasite biology, methods to induce potent cell-mediated immune responses, the difficulties associated with defining immune correlates of protection and antigen production and delivery. Finally, the use of attenuated organism vaccines is discussed.

This study determined whether the timing of re-feeding of protein-deficient mice restored functional protection against the gastrointestinal nematode, Heligmosomoides bakeri. Balb/c mice were fed a 3% protein-deficient (PD) diet and then transferred to 24% protein-sufficient (PS) diet either on the day of primary infection, 10 days after the primary infection, on the day of challenge infection, or 7 days after the challenge infection. Control mice were fed either the PD or PS diet. Onset of challenge, but not primary, infection caused short-term body weight loss, anorexia and reduced feed efficiency. Weight gain was delayed in mice when re-feeding commenced on the day of challenge infection; alkaline phosphatase (ALP) was also elevated in these mice on day 28 post-challenge. In contrast, other re-feeding groups attained similar body weights to PS mice within 4 days and had similar ALP at day 28. Serum leptin was higher in PD than PS mice and positively associated with food intake. As expected, worm survival was prolonged in mice fed the PD diet. However, egg production and worm burdens were similar in all re-feeding groups to the PS mice, indicating that protein re-feeding during either the primary or challenge infection rapidly restored normal parasite clearance.

Changes in gene expression in Biomphalaria glabrata following infection with Schistosoma mansoni have been investigated using a modified differential display approach. RNA was extracted from ovotestis, mantle tissue and anterior nephridium of control and exposed snails at 2 time-points (4 h and 24 h) post-exposure and analysed by RNA fingerprinting. A number of transcripts were identified; some novel and some homologous to mRNAs in GenEMBL that were previously unknown in B. glabrata. Down regulation of one 241 bp mRNA expressed sequence fragment – with an open reading frame showing 48% identity to a cytochrome p450 over 80 residues – has been confirmed using semi-quantitative RT-PCR. Preliminary classification of B. glabrata cyp450 sequence shows it to fall into CLAN 2 of the cytochrome p450 superfamily. Differential display has been successful in identifying changes in gene expression in Biomphalaria glabrata upon infection with Schistosoma mansoni and promises to be a useful technique for the investigation of the interaction between host and parasite.

The Monogenea fauna was studied from the gills of the catfish Pimelodus maculatus. Fish were caught from Río de la Plata, in Buenos Aires harbour (Argentina). Five species of Monogenea were found. The structure of the monogenean community was analysed at the component and infracommunity level. The nature of the monogenean community, its possible interactivity, seasonal variation and predictability are also discussed.

In 1922 J. W. W. Stephens the morphological characters of the asexual forms of a malaria parasite which he had studied in the blood of a patient from East Africa were different from those of any usually accepted species, and he proposed to establish the parasite as a new species with the name Plasmodium ovale. In 1927, in collaboration with D. U. Owen, he confirmed and extended the description from the study of a case infected in Nigeria. In 1930 Warrington Yorke and D. U. Owen studied another case from Nigeria and, by utilising this patient's blood in the practice of malariatherapy, they were able to show that the peculiar morphological features of the asexual stages of the parasite remain constant when it is passed from person to person by direct blood inoculation.

The investigation deals in detail with the structure of the immovable and movable parts of the head of the Nycteribiidae. The muscles of the head are also studied.

The head capsule is modified from that of the Hippoboscidae. The vertex is not subdivided into several anatomically differing subregions. The occiput is represented by the border of the occipital foramen only. The postgenae are separated by the median membranous region of the head, as in the Hippoboscidae. The ptilinum serves as a flexible joint connecting the fronto-clypeus with the vertex and the genae. The fronto-clypeus is much more modified than that of the Hippoboscidae and is movable. The eyes are reduced to one or two facets, or are entirely absent. The greatly modified ocular sclerites are present in Cyclopodia only.

The antennae consist of the pedicle and the flagellum, the latter being concealed in the cavity of the former. They have the same structure as those of the Hippoboscidae parasitising birds.

The proboscis has much in common with that of the Hippoboscidae though it differs greatly in detail. It is composed of the rostrum, the haustellum and the labella.

The rostrum belongs morphologically to the head and is formed of the same parts as in the Hippoboscidae. All the parts of the rostrum, with the exception of the tormae, have the same arrangement as in that family. The tormae are separate and are attached at the lateral borders of the fronto-clypeus.

The haustellum is composed of the theca, the labial gutter, the labrumepipharynx and the hypopharynx. The two former parts belong morphologically to the labium, but differ in detail from those of the Hippoboscidae. The very short labrum-epipharynx is enclosed in the posterior part of the labial gutter.

The hypopharynx is very slender and as long as the theca; it is enclosed in the cavity formed by the dorso-lateral parts of the labial gutter.

Labial glands are present in the bulbous part of the theca; their ducts open into the food canal. All the muscles of the bulbous part of the haustellum are homologous with those of the same part of the proboscis of the Hippoboscidae.

The labella are entirely membranous, are armed with two inner and six prestomal teeth and their surfaces have serrated ridges. There are six gustatory sensillae on the membrane of each labellum, which are connected by the intralabial canaliculi with the sense cells of the bulbous part of the theca.

The cutting action of the teeth of the labella depends upon the protractions and retractions of the narrow part of the theca. When this part retracts it everts the inner membranous walls of the labella, and the teeth, being attached to these walls, perform protrusible divergent movements.

NK cells represent one of the first lines of defence in the immune reaction after invasion of Leishmania parasites. Depletion of mouse natural killer (NK) cells dramatically enhances susceptibility of normally resistant mice. In this study we evaluated the fate of NK cells and parasites after contact formation. The hydrophilic fluorescent dye CMFDA (chloro-methylfluorescin diacetate) that allows analysis of cytotoxicity in flow cytometry and microscopy was used. Furthermore, these findings were confirmed with scanning and transmission electron microscopy. Direct contact points were found between Leishmania promastigotes and naïve human NK cells. These contacts were associated with transfer of cytosol by membrane bridges and cytotoxicity of NK cells against Leishmania. However, in contrast to other target cells which allow repeated exocytosis of lytic granules, contact with Leishmania causes immediate destruction of NK cells in a non-apoptotic way. Our results give a reasonable explanation for ex vivo observations of reduced NK cell numbers and impaired NK response in patients with acute cutaneous leishmaniasis. Animal models have clearly shown that NK cells play a key role in the induction and direction of the immune response. Thus inhibition of NK cells at the onset of infection would be advantageous for the survival of the parasite.

Although there has been a great deal of research effort within the last two decades on identifying the active components of the saliva of blood-sucking ticks, mosquitoes, biting flies, fleas and bugs, essentially neglected have been the human lice. Despite initial reports in the early part of this century suggestive of vasodilatory, anticoagulant and immunosuppressive properties of the saliva, for the next 50 years there were no biochemical studies on the active principles. Very recently, anatomical and biochemical studies have begun to characterize the bioactive molecules in lice saliva. The louse stocks a salivary vasodilator in excess over what is needed for a single bite, and injects similar amounts at each successive bite. The vasodilator in lice saliva appears to have different pharmacological properties than peroxidative, oxidative and maxidilan types of vasodilators reported from other blood-sucking insects. Possible anticoagulant activities have also been characterized. This belated, but welcome, interest comes at a time of resurgence of lice-born disease in certain parts of Africa, and of resistance to chemical control in Europe and North America.

A strain of E. tenella which had been passaged serially in 42 batches of chick embryos appeared to have lost its ability to produce the characteristically large second-generation schizonts in both the embryonic chorioallantois and in the chicken caeca. Numerous small schizonts (10–16 µm in diameter), which contained 8–30 small merozoites, replaced the large second-generation schizonts characteristic of this species. The small schizonts were numerous 3–5 days after inoculation and were restricted to epithelial cells of the embryo allantois when sporozoites were inoculated via the allantoic cavity. These schizonts also occurred in epithelial cells of the caeca of chickens given oocysts via the crop. Cell reactions in the chicken caeca in response to infection were minimal. Numerous minute lesions were present in the chorioallantois 4–6 days after inoculation; these were associated with the small schizonts and later with gamonts and oocysts. The strain reproduced well in the embryo but failed to induce gross pathogenic effects and no mortality resulted from the infection.

Only two passages of this strain in chickens were needed to restore a high degree of pathogenicity. This change was accompanied by the reappearance of the characteristically large second-generation schizonts in the tunica propria of the caecal wall.

Metabolomics analysis, which aims at the systematic identification and quantification of all metabolites in biological systems, is emerging as a powerful new tool to identify biomarkers of disease, report on cellular responses to environmental perturbation, and to identify the targets of drugs. Here we discuss recent developments in metabolomic analysis, from the perspective of trypanosome research, highlighting remaining challenges and the most promising areas for future research.

Toxoplasma gondii and Neospora caninum are closely related protozoan parasites which cause lowered production and increased abortion in dairy cows. The aim of the present study was to determine the seroprevalence of T. gondii and N. caninum infection in dairy cows in the Guangxi Zhuang Autonomous Region (GZAR), subtropical southern China. In total, 875 serum samples were collected from the tail veins of dairy cows in 6 main dairy cow-rearing districts of 4 administrative cities in GZAR. The samples were surveyed for T. gondii antibody using the Indirect Haemagglutination Test (IHA), and 365 of the serum samples were examined for N. caninum antibody by indirect Enzyme-Linked Immunosorbent Assay (ELISA). The overall seroprevalence of T. gondii in dairy cows was 13·71% (120/875), and the average seroprevalence of N. caninum was 15·07% (55/365). There were significant differences in the seroprevalence of N. caninum infection between different districts (P = 0·002, χ2 = 9·261). The highest prevalences of T. gondii and N. caninum were found in cows older than 8 years and those that had completed 5–6 pregnancies. Five cows (1·37%) presented antibodies against both T. gondii and N. caninum, and dairy cows with both T. gondii and N. caninum antibodies had higher abortion rates. The present results indicate widespread exposure of dairy cows to T. gondii and N. caninum in GZAR, subtropical southern China.

Difficulties encountered in the determination of species of Contracaecum from seals led the writer to undertake a comparative morpholgical study of a considerable amount of material. This included the Ascarids from seals collected by the Scottish National Antarctic Expedition (Scotia) in 1902–4, and deposited in the Royal Scottish Museum, Edinburgh, which were obtained on loan through the courtesy of Dr A. C. Stephen. In the present paper some results of this investigation are presented, and short notes are added on other species of Ascaridae recorded from Pinnipedia.

Non-splenectomized mice developed acute infections of Eperythrozoon coccoides after inoculation, but were then immune to re-infection. Splenectomy evoked acute infections that were heavier and longer in duration than the primary infection; during this phase, the parasites doubled in number in about 5 hr. The presence of splenic tissue transplanted subcutaneously did not prevent this evocation by splenectomy.

The number of eperythrozoa decreased rapidly after the peak of infection had been reached, but no immune factor was detected in the blood. No immune factor or infection was transmitted from mother to offspring through the milk.

Citrated blood remained infective for 11 days at 3° C., for 17 hr. at 16–17·7°C. and for 3 hr. at 37° C. It was not infective after 24 hr. at either 24° C. or 16–17·7°C., or after 5 hr. at 37° C. Parasitized blood was not infective after drying.

Citrated blood was infective to mice when given by mouth, but was not infective when applied externally. Urine and faeces were non-infective.

Eperythrozoa remained infective when infected blood was diluted with twice its volume of water and left for 24 hr. at 3° C. Phenol, 0·5%, rendered citrated blood non-infective in 1 hr. Neoarsphenamine, 0·1%, reduced the infectivity of citrated blood in 16 hr., and rendered it non-infective in 26 hr.

Cortisone did not affect the course of infection of E. coccoides in mice when given subcutaneously, 1·0 mg. per 20 g. mouse, once daily for 6–9 days.

Concurrent infection with Plasmodium berghei evoked relapses of E. coccoides during the terminal stages of the P. berghei infection.

A dark-ground photographic technique was used to analyse the reactions of Schistosoma mansoni miracidia to homogeneous solutions of snail–conditioned water (SCW). The most significant effect of this water was to increase miracidial turning. This effect was maintained under both acid and alkaline conditions, after passage of the SCW through a mixed bed resin and after chelation of either calcium or both calcium and magnesium ions. The stimulant in the water was unaffected by trypsin but was protease–sensitive, suggesting its possible identity as a peptide. The importance of ‘active spaces’ rather than concentration gradients in miracidial host–location was emphasized.

American tegumentary leishmaniasis (ATL) samples obtained from the lesions of patients with typical (n = 25, 29%), atypical (n = 60, 69%) or both (n = 2%) clinical manifestations were analysed by multilocus enzyme electrophoresis, hsp70 restriction-fragment length polymorphism (PCR-RFLP), hsp70 sequencing and phylogenetics methods. The hsp70 PCR-RFLP analysis revealed two different profiles whose the most samples differed from those expected for Leishmania braziliensis and the other Leishmania species tested: of 39 samples evaluated, two (5%) had a restriction profile corresponding to L. braziliensis, and 37 (95%) had a restriction profile corresponding to a variant pattern. A 1300-bp hsp70 gene fragment was sequenced to aid in parasite identification and a phylogenetic analysis was performed including 26 consensus sequences from the ATL patient's samples and comparing to other Leishmania and trypanosomatids species. The dendrogram allowed to observe a potential population structure of L. braziliensis complex in the studied region, emphasizing that the majority of clinical samples presented a variant genetic profile. Of interest, the L. braziliensis diversity was associated with different clinical manifestations whose parasites with hsp70 variant profile were associated with atypical lesions. The results may be helpful to improve the diagnosis, treatment and control measures of the ATL in endemic areas.

Toxoplasmosis is a worldwide zoonosis. However, data from Portugal are limited and a considerable part of the literature is in Portuguese. Currently, the rate of congenital infection in Portugal is unknown, and almost nothing is known of sequelae of congenital toxoplasmosis. There is no recent general population-based serological survey of Toxoplasma gondii in humans in Portugal. In addition, there is little information on genetic characteristics of T. gondii in animals and humans. In the present paper, we review prevalence, clinical spectrum and epidemiology of T. gondii in humans and animals in Portugal. This knowledge should be useful to biologists, public health workers, physicians and veterinarians.

Tuberculosis is an infectious disease associated with significant mortality and morbidity worldwide, particularly in developing countries. The rise of antibiotic resistance in Mycobacterium tuberculosis (Mtb) urgently demands the development of new drug leads to tackle resistant strains. Fragment-based methods have recently emerged at the forefront of pharmaceutical development as a means to generate more effective lead structures, via the identification of fragment molecules that form weak but high quality interactions with the target biomolecule and subsequent fragment optimization. This review highlights a number of novel inhibitors of Mtb targets that have been developed through fragment-based approaches in recent years.

Little is known about the types of intestinal parasites that infected people living in prehistoric Britain. The Late Bronze Age archaeological site of Must Farm was a pile-dwelling settlement located in a wetland, consisting of stilted timber structures constructed over a slow-moving freshwater channel. At excavation, sediment samples were collected from occupation deposits around the timber structures. Fifteen coprolites were also hand-recovered from the occupation deposits; four were identified as human and seven as canine, using fecal lipid biomarkers. Digital light microscopy was used to identify preserved helminth eggs in the sediment and coprolites. Eggs of fish tapeworm (Diphyllobothrium latum and Diphyllobothrium dendriticum), Echinostoma sp., giant kidney worm (Dioctophyma renale), probable pig whipworm (Trichuris suis) and Capillaria sp. were found. This is the earliest evidence for fish tapeworm, Echinostoma worm, Capillaria worm and the giant kidney worm so far identified in Britain. It appears that the wetland environment of the settlement contributed to establishing parasite diversity and put the inhabitants at risk of infection by helminth species spread by eating raw fish, frogs or molluscs that flourish in freshwater aquatic environments, conversely the wetland may also have protected them from infection by certain geohelminths.

The cloning and molecular characterization of a cDNA encoding Ascaris suum 24 kDa antigen (As24) are described. The cDNA sequence consists of 853 bp with an open reading frame coding for a protein of 147 amino acids with an inferred signal peptide of 19 amino acids. The predicted molecular mass and pI were 16 kDa and 8·35 respectively. The endogenous protein in adult A. suum was 24 kDa with the expected pI. A search of the public databases revealed over 50% homology with proteins from filarial parasites but not to other known proteins, suggesting that As24 is a nematode-specific protein. Immunohistochemical studies using polyclonal antibodies raised against Escherichia coli-expressed recombinant As24 demonstrated that the endogenous As24 proteins were intensely localized in unembryonated eggs within the uterus, uterine and gut epithelium, muscle tissues and in the hypodermis of an adult female A. suum. Endogenous As24 was expressed throughout A. suum development and was detected in the excretory/secretory products by immunoblot analysis. Importantly, a homologous protein(s) was detected in Ascaris from human and Toxocara canis from dog, suggesting that As24 is a nematode-specific protein.