Toxoplasma gondii, a zoonotic parasite of global importance, infects all endothermic vertebrates, with extensive health implications. The prevalence of this parasite is seldom monitored in wildlife. Here, a semi-aquatic species, the Eurasian otter (Lutra lutra) was used as a model to assess the potential effect of climate, land cover and biotic factors on T. gondii seroprevalence in British wildlife. The Sabin–Feldman cytoplasm-modifying dye test identified T. gondii antibodies in 25·5% of blood samples from otters found dead, mainly as road kill, in England and Wales, between 2004 and 2010. Otters in the east of England were more likely to be infected with T. gondii than those in western regions. Land cover and temperature are key determinants of T. gondii infection risk, with more infection in arable areas and lower infection where temperatures are higher. The probability of T. gondii infection increased with host age, reflecting cumulative exposure with time, but there was no association between T. gondii seroprevalence and cause of host death.

Besnoitia besnoiti and Toxoplasma gondii are two closely related parasites that interact with the host cell microtubule cytoskeleton during host cell invasion. Here we studied the relationship between the ability of these parasites to invade and to recruit the host cell centrosome and the Golgi apparatus. We observed that T. gondii recruits the host cell centrosome towards the parasitophorous vacuole (PV), whereas B. besnoiti does not. Notably, both parasites recruit the host Golgi apparatus to the PV but its organization is affected in different ways. We also investigated the impact of depleting and over-expressing the host centrosomal protein TBCCD1, involved in centrosome positioning and Golgi apparatus integrity, on the ability of these parasites to invade and replicate. Toxoplasma gondii replication rate decreases in cells over-expressing TBCCD1 but not in TBCCD1-depleted cells; while for B. besnoiti no differences were found. However, B. besnoiti promotes a reorganization of the Golgi ribbon previously fragmented by TBCCD1 depletion. These results suggest that successful establishment of PVs in the host cell requires modulation of the Golgi apparatus which probably involves modifications in microtubule cytoskeleton organization and dynamics. These differences in how T. gondii and B. besnoiti interact with their host cells may indicate different evolutionary paths.

Schistosoma japonicum, a human blood fluke, causes a parasitic disease affecting millions of people in Asia. Thioredoxin–glutathione system of S. japonicum plays a critical role in maintaining the redox balance in parasite, which is a potential target for development of novel antischistosomal agents. Here we cloned the gene of S. japonicum thioredoxin (SjTrx), expressed and purified the recombinant SjTrx in Escherichia coli. Functional assay shows that SjTrx catalyses the dithiothreitol (DTT) reduction of insulin disulphide bonds. The coupling assay of SjTrx with its endogenous reductase, thioredoxin glutathione reductase from S. japonicum (SjTGR), supports its biological function to maintain the redox homeostasis in the cell. Furthermore, the crystal structure of SjTrx in the oxidized state was determined at 2·0 Å resolution, revealing a typical architecture of thioredoxin fold. The structural information of SjTrx provides us important clues for understanding the maintenance function of redox homeostasis in S. japonicum and pathogenesis of this chronic disease.

Susceptibility to Echinococcus multilocularis infection considerably varies among intermediate (mostly rodents) and dead-end host species (e.g. humans and pig), in particular regarding intestinal oncosphere invasion and subsequent hepatic metacestode development. Wistar rats are highly resistant to infection and subsequent diseases upon oral inoculation with E. multilocularis eggs, however, after immunosuppressive treatment with dexamethasone, rats become susceptible. To address the role of the cellular innate immunity, Wistar rats were individually or combined depleted of natural killer (NK) cells, macrophages (MΦ) and granulocytes (polymorphonuclear cells, PMN) prior to E. multilocularis egg inoculation. Although NK cell and MΦ depletion did not alter the resistance status of rats, the majority of PMN-depleted animals developed liver metacestodes within 10 weeks, indicating that PMN are key players in preventing oncosphere migration and/or development in Wistar rats. In vitro studies indicated that resistance is not caused by neutrophil reactive oxygen species or NETosis. Also, light microscopical examinations of the small intestine showed that oral inoculation of E. multilocularis eggs does not elicit a mucosal neutrophil response, suggesting that the interaction of oncospheres and neutrophils may occur after the former have entered the peripheral blood. We suggest to consider granulocytes as mediators of resistance in more resistant species, such as humans.

Tams1 is the polymorphic immunodominant merozoite-piroplasm surface protein of Theileria annulata. Evidence for selection of divergent forms of Tams1 has been obtained recently. This study was performed to address whether selection takes place during persistent infection of the bovine host or during passage through the Hyalomma tick vector. Four calves were infected with a T. annulata isolate representing multiple parasite genotypes. The development of the parasite population was analysed by denaturing gradient gel electrophoresis (DGGE) using the Tams1 gene as a marker. In addition, the parasitaemia was measured by a semi-quantitative reverse line blot hybridization assay in order to correlate Tams1 variation to changes in parasitaemia. It was found that both parasitaemia and parasite population displayed limited variation during persistent infection. Ticks were allowed to acquire T. annulata during 2 periods of the bovine infection. Tams1 alleles detected in ticks fed during acute infection were identical to the population in the bovine host. However, ticks fed during the carrier status acquired parasites showing a single Tams1 isotype that represented, in several cases, a minor population in the bovine host at the time of infestation. Although only a limited number of ticks were studied, these preliminary data suggest that specific parasite genotypes may be selected during tick transmission from a carrier animal.

Dientamoeba fragilis is an inhabitant of human gastrointestinal tract with a worldwide distribution. The first description considered this protozoan a rare and harmless commensal, since then it has struggled to gain recognition as a pathogen. Commercial multiplex real-time PCR was used to detect D. fragilis in fecal samples from hospitalized children (⩽18 years) with acute gastrointestinal disease, admitted to two hospitals of Lisbon area, with different demographic characteristics. A total of 176 children were studied, 103 (58·5%) male, 144 (81·8%) children between 0 and 5 years and 32 (18·2%) above 6 years old. The overall protozoa frequency considering the four tested microorganisms were 8·5% (15/176), and the most frequently found protozoan was D. fragilis, 6·3% (11/176). Dientamoeba fragilis frequency was higher among older children (21·9%), than younger children (2·8%), and greater in boys (6·8%) than in girls (5·5%). All positive children presented with diarrhoea associated with vomiting, fever and abdominal pain. Infection was associated with the age of children (P < 0·001), school attendance (P = 0·002) and consumption of certain foods (P = 0·014), e.g. cakes with crème and ham. The frequency of diantamoebiasis found in a cohort of hospitalized Portuguese children, with acute gastrointestinal disease, could be considered a very high value when compared with the protozoan frequency normally associated with this pathology.

1. Progressive anaemia was observed in 30 White Leghorn hens obtained from a poultry farm in England, where the ailment became noticeable through the yellowish-paleness of the combs and gills of the diseased hens and the heavy mortality following on the appearance of these symptoms.

Only hens of one stock were affected, other breeds on the same farm did not develop the same or a similar condition.

2. The anaemia was characterised by a lasting diminution in the normal number of erythrocytes, a Hb index less than 1, the preponderance of immature and deformed erythrocytes and the presence of marrow-cells of the erythrocyte series in the circulating blood. The leucocytes were normal or slightly below normal in numbers; the monocytes were diminished.

3. The essential pathological lesion is an intravascular proliferation in the marrow of the stem cell of erythrocytes, which results in the blood-stream being overrun with erythroblasts and immature erythrocytes; these are rapidly destroyed in the spleen and liver, whilst the issue of normal erythrocytes in the marrow is hindered. The myeloblastic elements do not proliferate actively. These lesions explain the pathogenesis of the anaemia.

4. This intensified activity of the bone-marrow, the resulting congestion and increased localised pressure, induce the production of osteoid tissue; the femur and other bones of the fowls thicken, so that the medullary canal is nearly obliterated.

5. Attempts to detect microbes in the blood or isolate bacteria from the organs of anaemic hens failed. The spontaneously diseased hens all harbour a minute cestode, Davainea proglottina, in the duodenum, where the helminth causes round-celled infiltration, petechiae and thickening of the intestinal peritoneum.

6. In Davainea-free fowls of the same and other breeds, the injection of blood and cellular emulsions of liver and spleen from erythromyelotic fowls caused a condition of hypercythaemia which lasted several months. This was related to congestion and hyperplasia of the red-blood-marrow and accompanied by considerable enlargement of the liver and spleen.

7. Typical erythromyelosis was reproduced in one hen of the same stock from the same farm, by the intravenous inoculation of blood and tissue from spontaneously diseased fowls. In other hens only a transient type of anaemia was obtained, corresponding to the experience of other observers, more particularly Bedson and Knight (1924).

The experimental results agree with the view that a proliferative micro-plasm is the specific cause of this condition.

8. It is suggested that the development of this outbreak is induced by the massive infestation of the duodenum with Davainea proglottina. This irritative cestode produces a receptive condition in the bone-marrow of the fowl, which allows the microplasm to unfold its pathogenic properties. The duodenal parasite therefore plays the part of a preparatory cause. The association of cestode and microplasm affords an example of a “compound” disease.

9. The microplasm is specific and multiplying solely in certain definite blood-marrow cells of Gallus domesticus. This proliferative action can be compared to that of the microbes of epitheliosis contagiosa and Rous fowl-sarcoma; the resulting cell-reaction is then similar to tumour formation in these and other tissues of the fowl.

10. Since this haemopathy in fowls is characterised by specific pathological features in the erythrocyte-producing cells of the marrow, the name erythromyelosis is suggested, to mark clearly the position of the principal cellular lesion.

11. This disease (erythromyelosis) has been distinguished from other ailments of fowls, in the course of which anaemia may also occur, e.g. avian tuberculosis with involvement of the marrow. The leucocythaemic type is differentiated by diagnosis of the predominant type of cell found in active multiplication in the blood and the position of the cell-proliferation in the marrow.

12. Bone-marrow tumours and neoplasms of the lymphatic system occur in fowls, without causing progressive alterations in the cellular contents of the blood. The perivascular aggregations of cells in different organs may also hypertrophy, without any cytological changes being noticeable in the blood, except diminution of the numbers of erythrocytes. Such lesions are therefore distinguishable from different forms of leucocythaemia, where a persistent and progressive increase of genetically related cells is noticeable in the bloodstream and is concurrent with certain diffuse lesions of the marrow.

13. The haemoblastic diseases of fowls, such as erythromyelosis, leueo-cythaemia, lymphocythaemia, differ from tumours of the blood-producing tissues, since the former are the result of a pathological disturbance of the mechanism of haemocytopoiesis, whilst the latter are the localised proliferations of cellular elements, which then may spread by a process identical to neoplastic metastasis. There is a distinction in mode and degree.

14. The many resemblances between erythromyelosis and certain blood diseases in man and animals may indicate that their pathogenesis. though not identical, is comparable. A similar analogy may be applicable to fowl-leuco-cythaemia and lymphocythaemia.

During trawling expeditions off the coast of Galway in 1926–1928 I examined 100 specimens of Solea vulgaris Quensel, of varying sizes, and found about 60 per cent. infected with the parasite herein described. Usually two to five parasites were found attached to an infected fish, but up to eight were discovered on a single host.

Sieboldiellina planariarum is a large astomatous ciliate parasitic in the gut of the freshwater Triclad Planaria torva. The writer's attention was first called to it at Cambridge in 1923 whilst demonstrating the structure of Planarians collected from ditches in the neighbourhood.

Nippostrongylus brasiliensis of either sex, 216 h old, were injected directly in the intestinal lumen of 10-week-old laparotomized male rats in the duodenum or near the ileocaecal valve and their position ascertained 24 h later. Males migrated anteriad farther when females were placed in the duodenum than when they were alone or when 32P tagged males were in the duodenum. Females migrated anteriad similarly when they were alone or when males were in the duodenum or when 32 P tagged females were injected in the duodenum. The findings are consistent with the idea that females attract males anteriad at a distance.

Immunological and structural changes in the thymus and pronephros of Cyprinus carpio infected with the blood fluke, Sanguinicola inermis for 30 days, and exposed to 0·5mg NH+4/l for 48 and 168h were investigated. Ultrastructural observations revealed cell disruption and highly vacuolated cytoplasm in the thymus. Of the cells that remained intact there was a significant increase in thrombocytes after 48h exposure to the pollutant. In addition, there was a decrease in lymphocytes following exposure to ammonia at both time-periods studied. In contrast the pronephros of fish exposed to the pollutant underwent relatively mild changes in cellular architecture although ammonia and time of exposure had significant effects on the proportions of several leucocyte types. A significant decrease in neutrophils, thrombocytes and lymphocytes occurred in fish exposed to the pollutant for 168h. Pronephric lymphocyte stimulation (cpm) by Con A and PWM increased in vitro, whereas the stimulation index was reduced in infected fish exposed to ammonia. Changes in the immune organs of S. inermis-infected carp treated with pollutant were both organ- and time-specific. The possible reasons for this are discussed and significance in relationship to parasitization assessed.

Trypanosoma cruzi trypomastigotes occur as a mixture of morphologically distinct forms in natural and experimental infections. We have been able to separate mixed populations into slender and broad forms using a continuous gradient composed of 12·5–25·0% hypaque in culture media. Previously, comparisons could only be made by using various strains of T. cruzi which show a predominance of either form. This separation procedure gave recoveries of 88–98% and had no effect on the viability of the parasites. Infectivity studies showed that slender forms were much more infectious to cultured muscle cells than were broad forms. However, studies with macrophages from CFW mice showed both forms to be equally capable of infection. It is concluded that the mechanism by which each form infects host cells is different; slender forms being capable of infecting both by penetration and by phagocytosis, while broad forms are only capable of infecting through phagocytosis.

Attempts were made to immunize goats by infection with large numbers of metacyclic trypanosomes of a clone of Trypanosoma vivax, followed by chemotherapy. Five groups of 6 goats each were infected intradermally with 5 different doses of cultured metacyclics of T. vivax, ranging from 102 to 106 trypanosomes/goat. Four weeks after infection, the goats were treated with 10 mg/kg diminazene aceturate (Berenil, Hoechst A.G.). Three weeks after treatment, 3 goats in each group were challenged intradermally with 104 homologous metacyclics derived from culture. The remaining 3 goats in each group were challenged by 20 tsetse infected with the homologous clone. Five out of 30 goats were resistant to homologous challenge; 4 of the goats that had been challenged with cultured parasites, and 1 that had been challenged by tsetse. In each group 1 goat was protected. Protection was therefore not apparently influenced by the number of trypanosomes used to establish the primary infection. In another experiment, 6 goats were each infected by feeding 100 tsetse on the goats for 15 consecutive days. Three weeks after infection the goats were treated with Berenil and 3 weeks later challenged by 20 tsetse infected with the homologous clone. Three out of the 6 goats resisted challenge. The susceptible goats in both experiments, however, showed a reduction in the peak of parasitaemia following challenge compared with both challenge controls and the initial infections. Lytic antibodies to cultured metacyclics of T. vivax were detected in goats that resisted challenge after a primary infection with cultured metacyclics, and in resistant and susceptible goats after a primary infection by tsetse. All infected goats produced lytic antibodies to live bloodstream forms, as well as antibodies to bloodstream form lysates (demonstrated by ELISA). It is suggested that the immunity that had been induced in some of the experimental animals is due to antibody responses to both metacyclic and bloodstream variable antigen types (VATs) expressed during infection.

Fifty tetrathyridia of Mesocestoides corti showing various degrees of scolex fission during asexual multiplication were subjected to estimation of the total number of neurons in the central ganglia, using a ‘fractionator’ designed sampling scheme on every 10th Historesin section through the specimen. Neurons were stained with Cresyl fast violet and viewed by transmission light microscopy. Cells were only counted if a nucleolus was present. The animals were divided into groups according to their stage in the asexual reproduction process indicated by head shape and sucker number. Animals which had a narrow head with 4 suckers contained about 250 neurons. As their heads became wider, in preparation for fission, neurons were overproduced by up to 5-fold. As additional suckers were developed (8 in total), neurons were reduced to about 500. Finally, when the animals had 2 clearly discernible heads, with 4 suckers on each, the total number of neurons (per head) was again approximately 250. As the heads became wider again, neuron over-production recommenced. Exceptionally shaped animals with an X- or T-shaped single head were observed. These results suggest that there is a preparatory stage for neuronal regeneration, during which the basic population of neurons is about 250. This is followed by neuron over-production followed by a selective cell death of neurons which occurred during asexual multiplication. Another cycle for neuronal regeneration can occur before an animal completes its fission process.

The actual commencement of the great wave of human trypanosomiasis, which claimed so many thousands of lives along the shores of Lake Victoria, is difficult to determine. The attention of Europeans was first attracted to the disease early in 1901, when eight cases of sleeping sickness were admitted to Mengo hospital by the Drs Cook, who had been established in Uganda some five years, to whom the clinical picture was quite new. Enquiry among the chiefs of Buganda revealed the fact that sleeping sickness was known to the natives as mongota, and was more prevalent in the eastern portion of the Protectorate than in Buganda itself. A large number of deaths had already occurred in Busoga Province and in Buvuma Island.

Although immunisation protocols for a wide variety of parasitic diseases have been developed, it is often questioned why these do not always reach the market. In this review information about the regulations and procedures that apply to licensing the production and marketing of medicinal preparations, especially parasite vaccines, is presented. These general regulations specify issues on product (quality, safety, efficacy and potency) and production (facilities and consistency). Vaccine developers and manufacturers have to comply with these regulations, which may involve years of research and development. Moreover, where the manufacturer claims specific features of the product, these claims have to be corroborated by (experimental) data. A series of principles has been used to develop vaccines against parasite infections varying from the use of (attenuated) live vaccines to killed vaccines and subunit vaccines. The implications of some specific regulatory issues associated with these approaches are discussed.

Toxoplasma gondii is an obligate intracellular parasite that interacts with calcium storage organelles and induces calcium-dependent signalling in macrophages. This study was performed to determine whether Toxoplasma induces changes in intracellular calcium in these cells. Ratiometric imaging of live, Fura-2 loaded macrophages challenged with T. gondii revealed robust elevations in intracellular calcium. These elevations were late in onset, beginning 15–20 min after addition of parasites and occurred in up to 20% of macrophages in an imaging field. Further characterization of these events revealed that they follow from challenge with live T. gondii, but not heat-killed parasites or soluble Toxoplasma antigen (STAg). Parasite-induced calcium elevations derived from extracellular sources, and were independent of host recognition factors MyD88 and CCR5. These findings indicate that Toxoplasma gondii alters calcium homeostasis in macrophages and this activity is independent of known pathways involved in the innate recognition of this organism.

We studied the energy cost of egg production in two flea species (Parapulex chephrenis and Xenopsylla ramesis) feeding on principal (Acomys cahirinus and Meriones crassus, respectively) and auxiliary (M. crassus and A. cahirinus, respectively) rodent hosts. We predicted that fleas feeding on principal as compared with auxiliary hosts will (a) expend less energy for egg production; (b) produce larger eggs and (c) live longer after oviposition. Both fleas produced more eggs and spent less energy per egg when exploiting principal hosts. Parapulex chephrenis produced larger eggs after exploiting auxiliary hosts, while the opposite was true for X. ramesis. After oviposition, P. chephrenis fed on the auxiliary hosts survived for a shorter time than those fed on the principal hosts, while in X. ramesis the survival time did not differ among hosts. Our results suggested that one of the proximate causes for lower reproductive performance and subsequent lower abundance of fleas on auxiliary hosts is the higher energy cost of egg production. However, in some species, lower offspring number may be compensated to some extent by their size, although this compensation may also compromise their future reproduction via decreased survival. In addition, the reproductive strategy of exploitation of low profitable (i.e. auxiliary) hosts may differ between flea species.