The performance of 4 serological tests for the diagnosis of Chagas disease was evaluated in Santander, Colombia, a region still presenting active transmission. Serum samples from 638 individuals were submitted to an enzyme immunoassay test (EIA), using total lysate of a local Trypanosoma cruzi strain and 52·5% were positive (335/638). A subset of this group (94 positive individuals and 90 seronegatives) was randomly selected for further serological confirmation. Three additional tests were used – indirect immunofluorescence (IIF) and 2 distinct enzyme-linked immunosorbent assays using total lysate of the Y strain (EIA BM) and a mixture of 2 recombinant antigens (EIA RA). Seventy-nine patients were seropositive in all tests (84·0% – 79/94). The number of positive sera with the IIF, EIA RA and EIA BM was 84/94 (89·4%), 80/94 (85·1%) and 79/94 (84·0%), respectively. In 15 out of the 94 EIA seropositive patients (16·0%), 10 individuals were negative in all 3 tests (10·6% – 10/94). One was negative in the EIA BM and positive in the other two tests (1·1% – 1/94) and 4 patients were positive, solely, in the IIF assay (4·3% – 4/94). Relative to the 90 EIA negative individuals, 89 were confirmed in all other tests (98·9% – 89/90). One individual, although seronegative in the IIF, was positive in both confirmatory EIA tests (1·1% – 1/90). In addition, 120 blood specimens were submitted to PCR amplification. This group consisted of 79 confirmed seropositive cases, 16 individuals with discordant serological results and 25 validated seronegative individuals. The PCR was able to detect the presence of parasite DNA in 67 out of the 79 seropositive patients (84·8%), in 8 individuals with discordant serology (50·0%) and in only one seronegative individual (4·0%). The results pointed to the necessity for performing more than one serological test, preferentially with antigens from autochthonous strains, to achieve a reliable diagnosis of Chagas disease in Colombia.

The proportion of young honeybees which became infested with Acarapis woodi Rennie rose with an increased degree of infestation of the older bees and increased temperatures. But the most important influence controlling infestation seems to be the frequency with which young and old bees come into contact; this is probably determined by the foraging activity of the bees and the ratio of young bees to old bees; the percentage of infested bees in colonies diminished rapidly during nectar-flows and rose in periods of little nectar-flow.

The mortality of infested bees was only slightly greater than that of non-infested bees and became evident only after the bees had been in prolonged unfavourable circumstances. There were no obvious signs of disease in heavily infested colonies in summer. Colonies with more than about 30 or 40 % of infested bees seemed more likely to die in winter than non-infested colonies, but few colonies became so heavily infested even after a very poor season.

The fine structure of the flame cells, tubules and excretory pore of the miracidium of Fasciola hepatica are described. The tubules are formed from the flattened surfaces of cells rolled round and joined at their tips by a desmosome. Vesicles are present in the wall of the tubule, possibly indicating secretory activity. Clubshaped microvilli are present only in the terminal region near the excretory pore. Possible functions of the system are discussed.

The present study examines the role of host genetics in predisposition to Ascaris lumbricoides and Trichuris trichiura infection, by comparing the associations between age-standardized infection intensities of parents and their children (genetically related), with age-standardized infection intensities of parents alone (not related), within families previously demonstrated to exhibit familial predisposition. The lack of a consistent trend in infection intensity associations within families, in particular the lack of a stronger association between parents and their children than between unrelated parents, suggests that host genetic factors are not a major determinant of infection status. If there is a genetic basis for predisposition, then the data suggest that the effects of this genetic basis are overwhelmed by other, environmental or behavioural features of the family household.

The hatching mechanism of the liver fluke egg was investigated and a new interpretation put forward. Light stimulates the miracidium and, when fully activated, the miracidium is responsible for altering the permeability of the membrane on the internal concave surface of the viscous cushion. Prior to expansion the cushion (a fibrillar mucoprotein complex) is in a dehydrated or semihydrated state. The change in permeability of the enclosing membrane allows the fluid egg contents to permeate into the cushion. The fibrillar matrix becomes hydrated and an increase in volume occurs. This compresses the miracidium and sacs and an internal pressure is set up which gradually increases until the operculum finally ruptures and flies back violently. The cushion and miracidium are then expelled by the hypertonicity of the egg contents.

I would like to thank Professor O. W. Richards, in whose department this work was carried out, also Dr F. Call and Professor B. G. Peters for their many helpful criticisms. I am also grateful to Miss Mary Morris for assistance with the electron microscopy.

The migration of schistosomula through the lungs of mice has been investigated at the ultrastructural level. On Days 4 and 7 post-infection schistosomula were located only within the vasculature. Newly arrived Day 4 schistosomula were highly convoluted and completely occluded vessels measuring approximately 15–25μm in diameter. By Day 7 parasites were located in pulmonary capillaries, often causing considerable distension. The subtegumentary fibrous interstitial layer was absent in elongated lung schistosomula. The minimum diameter of such parasites was 8μm, and their tegument was usually unridged and flattened against the vascular endothelium, thus resulting in close apposition of endothelial and tegumental membranes. The tight fit of schistosomula in the pulmonary capillaries implies that migration through them is both a strenuous and slow process. The migratory delay in the lungs after arrival from the skin occurs because dimensional changes are necessary for successful capillary migration.

Factors which affect adhesion of cultured Trypanosoma congolense bloodstream forms to mammalian feeder cells have been examined. Using an in vitro binding assay, the initial events following interaction of trypanosomes with bovine aorta endothelial (BAE) cells were monitored by both light- and electron microscopy. Metabolic inhibitors and other biochemicals were incubated with either cells or parasites, to test whether any inhibited the process. Our findings suggest that adhesion of the parasites is an active process requiring metabolic energy from the trypanosomes, but not from endothelial cells. We also provide data suggesting that T. congolense bloodstream forms possess a lectin-like domain, localized at distinct sites on their flagellar surface, which interacts with specific carbohydrate receptors, most likely sialic acid residues, on the endothelial cell plasma membrane. We also suggest that the cytoskeletal protein actin is probably involved in this interaction.

The acquired immunity of twenty-eight splenectomized and twenty-three intact cattle following primary inoculation with either of two strains (the Y and the R strain) of Babesia bigemina was investigated. Some of these animals were carriers, while others had ceased to be infected. Observations were made on the response of splenectomized animals challenged up to four times with the original infecting (homologous) strain and finally with the heterologous strain.

In the splenectomized animals, a total of sixty-three homologous challenges produced parasitaemias in only twenty-seven instances, and these were generally delayed and of lower intensity than in control animals. Fifteen negative transmission tests performed shortly after thirty-two challenges showed that it was difficult to re-establish the homologous strain in some animals that had achieved self-cure. Substantial homologous immunity had developed within 16 weeks of the primary infection in animals which were no longer carrying B. bigemina.

All twenty-one splenectomized animals subjected to heterologous challenge developed well-marked parasitaemias. Nine of these had been highly immune to homologous challenge 2 weeks earlier. Eight of the twenty-one were carriers at the time of challenge and these showed much lower parasitaemias than the self-cured animals which almost invariably were acutely affected.

Challenges of twenty-three intact steers confirmed that the two strains differed immunologically. Six of twelve challenged homologously failed to show detectable parasitaemia, whereas all eleven challenged heterologously developed parasitaemias.

Killing of metacestodes by normal or post-infection macrophages and the regulation of this activity by cytokines were studied in vitro. The protoscolecidal activity of normal macrophages against Echinococcus granulosus was inhibited by a product of naive T-enriched lymphocytes co-cultured with protoscoleces (PSC). By contrast, supernates from co-cultures of Mesocestoides corti tetrathyridia (MCT) and T-enriched or B-enriched normal lymphocytes increased killing of MCT by normal macrophages. Larvicidal activity (against both PSC and MCT) was enhanced by high concentrations of macrophage-activating factors produced by Con A-stimulated rat lymphocytes (Con A-LK), but was reduced by low concentrations of these factors.

Until recently, almost all studies of leishmaniasis epidemiology were qualitative and descriptive. But now that the natural history of many Leishmania parasites is quite well known, there is growing interest in quantitative analysis. In this paper I use mathematical models in conjunction with field data to try to answer a wider range of questions than has previously been possible with descriptive techniques, and to sharpen some of the outstanding questions for laboratory workers. This is done with reference to the persistence and resilience of canine leishmaniasis, the maintenance of virulence poly morphisms in Leishmania populations, and the possible existence of cycles of human kala-azar. I conclude by posing a set of problems under three headings: diagnosis of infection (as distinct from disease), natural immunity to Leishmania infection in the vertebrate host, and genetic variation in the parasite population. Some solutions from the laboratory can be found in the companion paper by Black (1992).

1. A thirty-two-fold increase in resistance to sulphadiazine has been induced in Plasmodium gallinaceum in chicks by treatment with that drug.

2. No loss in resistance to sulphadiazine occurred in the resistant strain during cyclical passage through Aëdes aegypti.

3. The sulphadiazine-resistant strain was resistant also to sulphathiazole, sulphanilamide and sulphapyridine, but not to mepacrine, quinine or pamaquin. An increase in sensitivity to pamaquin was observed.

4. The sulphadiazine-resistant strain was resistant to paludrine and its methyl homologue M 4430.

5. In strains maintained in a state of acute infection and treated with sulphadiazine, resistance to paludrine developed more rapidly than resistance to sulphadiazine, and in one strain a high degree of resistance to paludrine was obtained before any increase in resistance to sulphadiazine could be detected.

6. Resistance to paludrine as induced by sulphadiazine, develops rapidly and extends at once over the full range of doses which the chick will tolerate, whereas resistance to paludrine as induced by that drug itself, develops more slowly and by stages.

7. Whereas resistance to paludrine is induced readily by treatment with sulphadiazine, resistance to sulphadiazine is induced by paludrine only after treatment with high doses of the drug for a prolonged period.

8. In latent infections of Plasmodium gallinaceum resistance to paludrine or sulphadiazine was not induced by sulphadiazine during the period of the experiment (49–75 days), though the aggregate dosage of drug used was much greater than that with which resistance was induced in strains maintained in an acute state.

9. It is not considered probable that cross-resistance between sulphadiazine and paludrine is due to a similar mode of action of these drugs, as whereas sulphadiazine is antagonized by p-amino-benzoic acid paludrine is not.

Maturity, mass and sex were determined for individual Raphidascaris acus from northern pike, Esox lucius from Dauphin Lake, Manitoba. Length, width and total fecundity of female worms; seasonal changes in worm numbers, growth and maturity; and inequalities in mass and fecundity were quantified for 8 sample periods between April and September. Recruitment of R. acus is most rapid in early spring and continues throughout summer. An accumulation of larvae over winter leads to a peak in the number of gravid worms in spring, while increased feeding by post-spawning pike on yellow perch, Perca fluviatilis, leads to lower levels of gravid worms throughout summer and early autumn. Gravid worms found in spring versus summer were similar with respect to all measured variables and do not appear to be from genetically distinct populations. The strength of the correlation between fecundity and mass for worms was variable between pike and between sample periods, and only weakly correlated when the data were pooled. Much of this variation could be attributed to continued growth of gravid worms after maturation. Worms found in June were smaller, less fecund and less variable and inequalities in mass and fecundity measured using Gini coefficients (GCs) and Lorenz curves (LCs) were most similar at this time. For all other sample periods inequality in fecundity was greater than the corresponding value for mass. Similarity between GCs based on mass and fecundity was maximum when the coefficient of variation (C.V.) for mass was greater than the corresponding C.V. for fecundity. These observations suggest that one must be cautious when applying GCs and LCs to the analysis of parasite populations.

The effects of parasitism by the daughter sporocyst of Cercaria bucephalopsis haimaena Lacaze-Duthiers, 1854, on the digestive cells of Cardium edule L. was investigated by comparing histochemically stained cells in healthy and parasitized animals. Two effects were apparent: (a) the branches of the daughter sporocysts compress the walls of the digestive gland ducts and tubules, and cut off the more distal tubule cells from their food supply, inducing starvation autolysis, (b) in digestive tubule cells not so cut off, glucose, glycogen, glycoproteins, phospholipid and proteolipid food storage globules and acid mucopolysaccharide are reduced. Neutral lipids, fatty acids, alkaline phosphatase, acid phosphatase and non-specific esterase are increased and there is also a compensatory increase in the number of food vacuoles. In the visceral haemocoel, in the neighbourhood of the parasite, glucose, fatty acids, neutral lipids and acid mucopolysaccharide are increased and glycogen reduced.

We are grateful to Professor E. W. Knight-Jones, for helpful comments and good working facilities, and to the Science Research Council, for the grant of a research studentships to one of us (E.A.B.).

Voltage-gated calcium (Ca2+) channels contribute to impulse propagation in excitable cells and also regulate intracellular levels of Ca2+. High voltage-activated (HVA) Ca2+ channels are heteromultimeric membrane proteins. The pore-forming, voltage-sensing subunit is the α1 subunit. We have cloned 3 HVA Ca2+ channel α1 subunit cDNAs from Schistosoma mansoni. One of these sequences most closely resembles the L-type class of HVA α1 subunits. The other two sequences are most closely related to non L-type α1 subunits. These schistosome α1 subunits have many of the features common to HVA Ca2+ channels, but also have distinct structural motifs. Analysis of the structural and functional properties of schistosome Ca2+ channel subunits may provide information about these critical components of excitable cells.

The aim of this investigation was to examine whether macrophage and Leishmania major glutathione were involved in either host or parasite protection against NO cytotoxicity. Buthionine sulfoximine (BSO), an inhibitor of γ-glutamyl-cysteine synthase, caused a complete and irreversible depletion of macrophage glutathione, but only a 20% and reversible decrease in L. major glutathione. Glutathione-depleted macrophages, when activated with IFN-γ/LPS, released less than 60% of the NO produced by untreated macrophages, resulting in a corresponding decrease in their leishmanicidal activity. BSO-treated macrophages were more susceptible to the cytotoxic effects of the NO donor SNAP. Treatment of macrophages with 1,3-bis(chloroethyl)-1-nitrosourea (BCNU), an inhibitor of glutathione reductase and trypanothione reductase or with Br-Octane, a glutathione-S-transferase substrate, resulted in a transient decrease in glutathione levels and did not increase the susceptibility of the macrophages to SNAP. Treatment of the promastigote forms of L. major with BCNU resulted in an 80% decrease in total glutathione concentration with no concomitant change in viability. However, this treatment rendered the parasites more susceptible to SNAP. Finally, macrophage glutathione protected the internalized L. major from SNAP. Overall, these results demonstrate that glutathione is an essential protective component against NO cytotoxicity on both macrophages and parasites.

Lymphatic filariasis (LF), which is highly endemic in 73 countries worldwide, is targeted for elimination by 2020. The strategy for achieving this goal is based on 4 sequential programmatic steps: mapping, Mass drug administration (MDA) implementation, post-MDA surveillance and verification of LF elimination. All 4 stages of the implementation process are dependent on the availability of user friendly and highly sensitive rapid diagnostic tools. By the end of 2012, 59 countries had completed mapping for LF and Eritrea was the only country yet to start the process. Rolling out new diagnostic tools to facilitate the mapping process will enable an accelerated shrinking of the LF map to zero endemic countries by 2020. When the Global Programme to Eliminate Lymphatic Filariasis was launched in 2000, diagnostic tools for LF were limited to clinical examination, detection of microfilaria (MF) by microscopy in night blood samples and detection of antibodies to native-antigen preparations. There has been a significant improvement in the traditional LF diagnostic methods in recent years and some new tools are now available. This paper provides an update on the human diagnostic tests available for LF and their current applications as tools in mapping and transmission monitoring. The values of entomological indicators and parasite detection and speciation methods applied to vector populations are also discussed.

African sleeping sickness is a parasitic disease transmitted through the bites of tsetse flies of the genus Glossina. We constructed mechanistic models for the basic reproduction number, R0, of Trypanosoma brucei gambiense and Trypanosoma brucei rhodesiense, respectively the causative agents of West and East African human sleeping sickness. We present global sensitivity analyses of these models that rank the importance of the biological parameters that may explain variation in R0, using parameter ranges based on literature, field data and expertize out of Uganda. For West African sleeping sickness, our results indicate that the proportion of bloodmeals taken from humans by Glossina fuscipes fuscipes is the most important factor, suggesting that differences in the exposure of humans to tsetse are fundamental to the distribution of T. b. gambiense. The second ranked parameter for T. b. gambiense and the highest ranked for T. b. rhodesiense was the proportion of Glossina refractory to infection. This finding underlines the possible implications of recent work showing that nutritionally stressed tsetse are more susceptible to trypanosome infection, and provides broad support for control strategies in development that are aimed at increasing refractoriness in tsetse flies. We note though that for T. b. rhodesiense the population parameters for tsetse – species composition, survival and abundance – were ranked almost as highly as the proportion refractory, and that the model assumed regular treatment of livestock with trypanocides as an established practice in the areas of Uganda experiencing East African sleeping sickness.

Parasites are considered to be an important selective force in host evolution but ecological studies of host-parasite systems are usually short-term providing only snap-shots of what may be dynamic systems. We have conducted four surveys of helminths of bank voles at three ecologically similar woodland sites in NE Poland, spaced over a period of 11 years, to assess the relative importance of temporal and spatial effects on helminth infracommunities. Some measures of infracommunity structure maintained relative stability: the rank order of prevalence and abundance of Heligmosomum mixtum, Heligmosomoides glareoli and Mastophorus muris changed little between the four surveys. Other measures changed markedly: dynamic changes were evident in Syphacia petrusewiczi which declined to local extinction, while the capillariid Aonchotheca annulosa first appeared in 2002 and then increased in prevalence and abundance over the remaining three surveys. Some species are therefore dynamic and both introductions and extinctions can be expected in ecological time. At higher taxonomic levels and for derived measures, year and host-age effects and their interactions with site are important. Our surveys emphasize that the site of capture is the major determinant of the species contributing to helminth community structure, providing some predictability in these systems.

In Europe, the zoonotic cycle of Babesia microti has not been determined so far. Recently, B. microti was detected in Ixodes ricinus ticks in Slovenia by using molecular methods. In order to investigate the mammalian hosts of B. microti in Slovenia we collected 261 small mammals representing 11 species. They were tested for the presence of babesial parasites with a PCR assay based on the nuclear small subunit rRNA gene (nss-rDNA). The bank vole (Clethrionomys glareolus) and yellow-necked mouse (Apodemus flavicollis) were infected with B. microti. The prevalence rate was 15·9% for C. glareolus and 11·8% for A. flavicollis. Nucleotide sequences of amplified portions of B. microti nss-rDNA from C. glareolus and A. flavicollis were indistinguishable from each other and identical with those previously described in I. ricinus ticks collected in Slovenia. The results of this study represent molecular evidence of B. microti in small mammals in Europe.

The oyster Crassostrea gasar is a species widely used as food and a source of income for the local population of the estuaries of Northeast Brazil. Perkinsus marinus and Perkinsus olseni are deleterious parasites for oyster farming and were recently detected in Brazil. In this study, a histopathologic survey of the oyster C. gasar cultured in the estuary of the River Mamanguape (Paraíba State) was performed. Adult oysters were collected in December 2011 and March, May, August and October 2012 and processed for histology and Perkinsus sp. identification by molecular analyses. Histopathological analysis revealed the presence of parasitic organisms including viral gametocytic hypertrophy, prokaryote-like colonies, protozoans (Perkinsus sp. and Nematopsis sp.) and metazoans (Tylocephalum sp. and cestodes). Other commensal organisms were also detected (the protozoan Ancistrocoma sp. and the turbellarian Urastoma sp.). The protozoan parasite Perkinsus sp. had the highest overall prevalence among the symbiotic organisms studied (48·9%), followed by Nematopsis sp. (36·3%). The other organisms were only sporadically observed. Only the protozoan Perkinsus sp. caused alterations in the oysters’ infected organs. Molecular analyses confirmed the presence of P. marinus, P. olseni and Perkinsus beihaiensis infecting the oyster C. gasar. This is the first report of P. beihaiensis in this oyster species.