Infections caused by protozoan parasites are among the most widespread and intractable transmissible diseases affecting the developing world, with malaria and leishmaniasis being the most costly in terms of morbidity and mortality. Although new drugs are urgently required against both diseases in the face of ever-rising resistance to frontline therapies, very few candidates passing through development pipelines possess a known and novel mode of action. Set in the context of drugs currently in use and under development, we present the evidence for N-myristoyltransferase (NMT), an enzyme that N-terminally lipidates a wide range of specific target proteins through post-translational modification, as a potential drug target in malaria and the leishmaniases. We discuss the limitations of current knowledge regarding the downstream targets of this enzyme in protozoa, and our recent progress towards potent cell-active NMT inhibitors against the most clinically-relevant species of parasite. Finally, we outline the next steps required in terms of both tools to understand N-myristoylation in protozoan parasites, and the generation of potential development candidates based on the output of our recently-reported high-throughput screens.

1. Tick paralysis occurs in man, the dog, and other domesticated animals on the East Coast of Australia.

2. As far as is known, the disease in Australia is only conveyed by the mature female of Ixodes holocyclus. The immature stages and the male of this species of ticks are non-pathogenic.

3. A similar disease, induced in a like manner by females of other species of ticks, has been observed by various authors in South Africa, Canada and Crete.

4. A single tick, as shown by experiment, may cause fatal paralysis in man and the dog in Australia, the death-rate in dogs being very high.

5. The main symptoms of tick paralysis in Australia do not differ materially from those observed in other countries. They begin with loss of coordination commencing in the hind legs in dogs, the fore legs, head and neck becoming subsequently affected. Death in all cases appears to be due to respiratory paralysis.

6. The onset of symptoms has not been observed to occur in less than 5 days from the time the tick or ticks have attached themselves to a dog.

7. Some of the ticks do not produce paralysis and others do, as shown repeatedly from my experiments on dogs. This requires explanation. It does not depend on the time that has elapsed between the tick having moulted and its attacking the dog.

8. Paralysis has never been seen to occur when ticks were removed from dogs before the time when symptoms usually appear.

9. It is impossible to transmit the disease from affected to healthy animals by the inoculation of blood, cerebro-spinal fluid or nervous tissue. Nor can the disease be transmitted by the injection of the body contents of ticks removed from affected animals. The examination of the fluids and organs of affected animals has failed to reveal the presence of pathogenic organisms.

10. It is considered that the causal factor in the production of the disease is a tick-derived toxin, this toxin being secreted by the salivary glands.

11. During the process of engorgement the salivary glands develop greatly, pouring out a maximum amount of secretion during the 2 days before the female tick drops.

12. An emulsion of salivary glands of I. holocyclus proved to be toxic for when injected into a dog (Experiment V, p. 424) it caused fever, vomiting, etc. The salivary secretion contains anticoagulins. As Nuttall and Strickland found for anticoagulins, individual ticks may vary in toxicity.

13. The period which elapses between the attachment of a tick to the host and the first signs of an attack of tick paralysis cannot be regarded in the light of an incubation period; it is dependent on the rate and stage of the tick's engorgement.

14. If it be accepted that the causal agent of the disease is a tick-derived toxin, it remains to be shown how such a toxin is produced.

15. There is some evidence that immunity to the disease may be acquired after recovering from previous attacks.

16. There is no evidence of season having an influence on tick paralysis. It was induced experimentally both in winter (June–August) and summer (November–December) in Sydney.

It is shown that under the conditions that prevailed in New Zealand in the late 1950s, Taenia hydatigena was hyperendemic, the life-cycle being regulated by a density-dependent constraint in the form of acquired immunity, and T. ovis was rare. The control measures that caused Echinococcus granulosus, which was endemic at the time, to decline towards extinction reduced T. hydatigena and T. ovis to endemic status only. A non-linear integrodifferential equation model, which was previously linearized to describe the life-cycle of E. granulosus in dogs and sheep in New Zealand, is used to describe the life-cycles of T. hydatigena and T. ovis. The model is then used to compare and contrast the population dynamics of these three species. The model is used to demonstrate that the endemic steady state is structurally unstable, and may be asymptotically unstable to small perturbations. It is also shown that despite the lower infection pressure experienced by the intermediate host in the endemic state, the numbers of larvae in sheep may be higher than in the hyperendemic state. Finally it is shown that the partial success of the control measures against T. hydatigena may have caused an increase in the numbers and prevalence of T. ovis larvae in sheep due to the reciprocal immunity between the two species.

Two distinct families of neuropeptides are known to endow platyhelminth nervous systems – the FMRFamide-like peptides (FLPs) and the neuropeptide Fs (NPFs). Flatworm FLPs are structurally simple, each 4–6 amino acids in length with a carboxy terminal aromatic-hydrophobic-Arg-Phe-amide motif. Thus far, four distinct flatworm FLPs have been characterized, with only one of these from a parasite. They have a widespread distribution within the central and peripheral nervous system of every flatworm examined, including neurones serving the attachment organs, the somatic musculature and the reproductive system. The only physiological role that has been identified for flatworm FLPs is myoexcitation. Flatworm NPFs are believed to be invertebrate homologues of the vertebrate neuropeptide Y (NPY) family of peptides. Flatworm NPFs are 36–39 amino acids in length and are characterized by a caboxy terminal GRPRFamide signature and conserved tyrosine residues at positions 10 and 17 from the carboxy terminal. Like FLPs, NPF occurs throughout flatworm nervous systems, although less is known about its biological role. While there is some evidence for a myoexcitatory action in cestodes and flukes, more compelling physiological data indicate that flatworm NPF inhibits cAMP levels in a manner that is characteristic of NPY action in vertebrates. The widespread expression of these neuropeptides in flatworm parasites highlights the potential of these signalling systems to yield new targets for novel anthelmintics. Although platyhelminth FLP and NPF receptors await identification, other molecules that play pivotal roles in neuropeptide signalling have been uncovered. These enzymes, involved in the biosynthesis and processing of flatworm neuropeptides, have recently been described and offer other distinct and attractive targets for therapeutic interference.

The global seroprevalence of latent toxoplasmosis is estimated to be higher than 30%. The presence of slowly dividing parasites in tissue cysts located mainly in immunoprivileged organs was long considered asymptomatic. Recently, many studies have shown that latent Toxoplasma infections could have serious impacts on human health. Here we ran a cross-sectional study in a population of 1486 volunteers. The results showed that 333 infected subjects scored worse than 1153 controls in 28 of 29 health-related variables. Similarly, they reported higher rates of 77 of a list of 134 disorders reported by at least 10 participants of the study. Toxoplasmosis was associated most strongly with musculoskeletal (τ = 0·107, P < 0·0005), followed by neurological (τ = 0·088, P < 0·0005), immune (τ = 0·085, p < 0·0005), metabolic (τ = 0·079, P < 0·0005), respiratory (τ = 0·068, P = 0·0001), allergic (τ = 0·053, P = 0·004), digestive system (τ = 0·052, P = 0·004) and mental health disorders (τ = 0·050, P = 0·008). Results of the present cohort study, along with the previous data from many case-control studies or ecological studies suggest that latent toxoplasmosis represents a large and so far underrated public health problem.

Surveys on Trichinella parasites in domestic and sylvatic animals collected in France, Italy, and in the Extremadura region of Spain showed that the distribution of Trichinella spiralis and Trichinella britovi is influenced by both environmental and human behaviour factors. In France, both Trichinella species are prevalent in the fox population from mountain areas and natural parks but are infrequent in wild boars (< 0·001%). In Italy, only T. britovi is present in sylvatic animals (foxes, wolves, and mustelids) living 500 m above sea level. This species is rare in wild boars ( < 0·001%) in that area. Sylvatic trichinellosis is found in only 24% and 34% of French and Italian territory, respectively, while lowland areas may generally be considered Trichinella-free, because the domestic cycle is absent. The ecology of T. spiralis and T. britovi in the Extremadura shows a different picture from that observed in France and Italy because of the presence of both domestic and sylvatic cycles. The domestic cycle not only allows the maintenance of T. spiralis in the domestic environment, but it also has a great impact on the prevalence in wild boar populations. It does not influence the prevalence in vulpine populations. These data suggest (1) that domestic trichinellosis occurs only in rural areas of Western Europe in association with traditional swine-rearing practices, but not in industrialized pig farms; (2) that sylvatic trichinellosis occurs only in natural habitats which, in Western Europe, are widespread in mountain areas; (3) that the fox is the primary reservoir in the sylvatic cycle, where the parasite is maintained in a closed circuit and (4) that among sylvatic animals T. spiralis is present at lower altitude than is T. britovi.

A new method has been established to define the limits on a spontaneous mutation rate for a gene in Plasmodium falciparum. The method combines mathematical modelling and large-scale in vitro culturing and calculates the difference in mutant frequencies at 2 separate time-points. We measured the mutation rate at 2 positions in the dihydrofolate reductase (DHFR) gene of 3D7, a pyrimethamine-sensitive line of P. falciparum. This line was re-cloned and an effectively large population was treated with a selective pyrimethamine concentration of 40 nM. We detected point mutations at codon-46 (TTA to TCA) and codon-108 (AGC to AAC), resulting in serine replacing leucine and asparagine replacing serine respectively in the corresponding gene product. The substitutions caused a decrease in pyrimethamine sensitivity. By mathematical modelling we determined that the mutation rate at a given position in DHFR was low and occurred at less than 2.5×10−9 mutations/DHFR gene/replication. This result has important implications for Plasmodium genetic diversity and anti-malarial drug therapy by demonstrating that even with low mutation rates anti-malarial resistance will inevitably arise when mutant alleles are selected under drug pressure.

This article presents an overview on phosphoglycan-containing components secreted by the insect and mammalian stages of several species of Leishmania, the causative agents of leishmaniasis in the Old and New World. Firstly, promastigotes of all three species considered, L. mexicana, L. donovani and L. major, shed lipophosphoglycan (LPG) into the culture medium possibly by release of micelles from the cell surface. Like the cell-associated LPG, culture supernatant LPG is arhphiphilic and composed of a lysoalkylphosphatidylinositol-phosphosaccharide core connected to species-specific phosphosaccharide repeats and oligosaccharide caps. Secondly, all three species release hydrophilic phosphoglycan. Thirdly, all three species appear to secrete proteins covalently modified by phosphosaccharide repeats and oligosaccharide caps. In the case of promastigotes of L. mexicana, these components are organized as two filamentous polymers released from the flagellar pocket: the secreted acid phosphatase (sAP) composed of a 100 kDa phosphoglycoprotein and a protein- containing high-molecular-weight-phosphoglycan (proteo-HMWPG) and fibrous networks likewise composed of phosphoglycan possibly linked to protein. Structural analyses and gene cloning suggest that the parasites can covalently modify protein regions rich in serine and threonine residues by the attachment of phosphosaccharide repeats capped by oligosaccharides. We propose that the networks formed in vitro correspond to fibrous material previously demonstrated in the digestive tract of infected sandflies. In the case of L. donovani, the sAP is also modified by phosphoglycans but contains neither proteo-HMWPG nor does it aggregate to filaments. Finally, L. mexicana amastigotes release proteo-HMWPG via the flagellar pocket into the parasitophorous vacuole of infected macrophages. This material appears to be released into the tissue of the infected mammal upon rupture of infected macrophages during lesion development. This secretory product may contribute to the pathology of lesion development.

1. Maturation of the plerocercoid of the pseudophyllidean cestode Diphyllobothrium sp. and the metacercaria of the strigeid trematode Diplostomum phoxini follow a similar pattern, which may be divided into the following phases: (1) cell multiplication; (2) segmentation or body shaping; (3) organogeny; (4) early gametogeny; (5) late gametogeny; (6) egg-shell formation and vitellogenesis; (7) oviposition.

2. Cytological or histochemical criteria have been developed by means of which each developmental phase may be recognized. Development in vitro may be considered comparable with that occurring in vivo if the same phases are completed with the same incubation time and without the appearance of cytological abnormalities.

3. The cell multiplication phase is characterized by high mitotic activity. This activity in vitro can be evaluated by incubating larvae after 24 hr. culture for a standard time (5 hr. Diphyllobothrium; 4 hr. Diplostomum) in 10−4 colchicine and comparing the number of mitoses seen in a defined area of an aceto-orcein squash with those found in tissue matured in vivo for the same period. Use of this criterion enables the growth-producing properties of a medium to be assessed after 24 hr. culture.

4. Phases 2–4 are characterized by the appearance of the tubular genitalia, the rudiments of which are readily recognized in aceto-orcein squashes or whole mount preparations.

5. The presence of spermatids and mature spermatozoa (phases 4–5) may be detected in aceto-orcein squashes or in fresh tissue teases. The quantity and activity of spermatozoa also serve as important criteria.

6. Recognition of phase 6, egg-shell formation, is carried out by means of diazo reagents which give orange or orange-red reactions with the phenolic egg-shell precursors in the vitellaria, as seen in whole mount preparations. Abnormal egg-shell development is reflected in the failure of vitellaria to give a normal diazo reaction.

7. The appearance of eggs which are macroscopically normal constitutes the final maturation phase. Early detection of eggs (especially abnormal ones) is facilitated by the use of the diazo reaction or the catechol test for polyphenol oxidase.

8. Embryonation and hatching of eggs to produce morphologically normal infective coracidia or miracidia serve as a final criteria.

9. Criteria for the maintenance of the adult condition are also considered.

Macrophage-like defence cells (haemocytes) of the pond snail Lymnaea stagnalis mediate cytotoxicity through reactive oxygen intermediates (ROIs). This activity is NADPH-oxidase dependent, as in mammalian phagocytes during the respiratory burst. In this study, mother sporocysts of schistosomes, the compatible Trichobilharzia ocellata and the incompatible Schistosoma mansoni evoke in vitro ROI activities (detected by luminol dependent chemiluminescence, LDCL) from L. stagnalis haemocytes. S. mansoni is encapsulated by haemocytes and eliminated, whereas T. ocellata escapes encapsulation and survives. Both schistosomes were equally susceptible to in vitro oxidative damage from exposure to hydrogen peroxide and to ROIs generated by a xanthine/xanthine oxidase system. Protocatechuic acid, a specific antagonist of NADPH-oxidase, delayed the killing of T. ocellata and S. mansoni sporocysts by haemocytes of resistant snails (Biomphalaria glabrata and L. stagnalis, respectively). We conclude that ROIs take part in haemocyte-mediated cytotoxicity. However, neither a snail's capability to generate ROIs, nor a schistosome's susceptibility to ROIs, determine snail/schistosome incompatibility. Snail/schistosome compatibility is rather determined by the parasite's ability modulate haemocyte behaviour such that effective encapsulation and the generation of lethal concentrations of ROIs are prevented.

Bovine besnoitiosis is caused by the cyst-forming apicomplexan parasite Besnoitia besnoiti. This disease progresses in two sequential phases: a febrile acute phase with oedemas and respiratory disorders, and a chronic phase characterized by the presence of subcutaneous tissue cysts and skin lesions. Serious consequences of the infection are poor body condition, sterility in bulls and eventual death. The role of host/parasite-dependent factors, which play a major role in the pathogenesis of the disease, is not yet fully elucidated. Isolate/strain virulence, parasite stage, dose and the route of parasite inoculation were studied under different experimental conditions, which make it difficult to compare the results. Data on host-dependent factors obtained from naturally infected cattle showed that (i) the seroprevalence of infection is similar in both sexes; (ii) seropositivity increases with age; (iii) both beef and dairy cattle are susceptible to the infection; and (iv) the cell-mediated immune response is likely to play a major role because a T cell response has been observed around several tissue cysts. Whether colostral antibodies are protective and to what extent the humoral immune response might reflect the disease/protection status require further research. Thus, a well-established experimental bovine model could help to clarify these important questions. The dynamics of B. besnoiti infection in cattle and available knowledge on relevant factors in the pathogenesis of the infection are reviewed in the present work.

Neuropeptides act as chemical signals in the nervous system to modulate behaviour. With the ongoing EST projects and DNA sequence determination of different genomes, the identification of neuropeptide genes has been made easier. Despite the relatively ‘simple’ repertoire of behaviours in the nematode Caenorhabditis elegans, this worm contains a surprisingly large and diverse set of neuropeptide genes. At least 109 genes encoding over 250 potential neuropeptides have been identified in C. elegans; all genes are likely to be expressed and many, if not all, of the predicted peptides are produced. The predicted peptides include: 38 insulin-like peptides, several of which are involved in development and reproductive growth, and over 70 FMRFamide-related peptides, some of which are involved in locomotion, reproduction, and social behaviour. Many of the C. elegans peptides are identical or highly similar to those isolated or predicted in parasitic nematodes, such as Ascaris suum, Haemonchus contortus, Ancylostoma caninum, Heterodera glycines and Meloidogyne arenaria, suggesting that the function of these peptides is similar across species. The challenge for the future is to determine the function of all the genes and individual peptides and to identify the receptors through which the peptides signal.

It has recently been shown using genetic markers that Ascaris in humans and pigs in Central America comprise reproductively isolated populations. We present a similar analysis for a region of China in which close association between pigs and humans has been the norm for thousands of years, and agricultural practices will result in frequent exposure to eggs from both sources. DNA fragments from selected regions of mitochondrial and ribosomal DNA were amplified by PCR and allelic forms identified following digestion with a panel of restriction enzymes, using DNA from a total of 115 individual worms from both people and pigs from 2 neighbouring villages. Significant frequency differences in both mtDNA haplotypes and the rDNA spacer were found between the 2 host-associated populations, indicating that they represented reproductively isolated populations. Mitochondrial haplotype frequencies were different from those observed in Guatemala and also from other Asian Ascaris populations, suggesting low levels of gene flow between populations. However, we found no evidence for significant heterogeneity in the genetic composition of Ascaris infrapopulations in either humans or pigs, possibly indicative of agricultural practices in China which have resulted in a random distribution of alleles within the parasite populations.

Studies on avian haemosporidia are on the rise, but we still lack a basic understanding of how ecological and evolutionary factors mold the distributions of haemosporidia among species in the same bird community. We studied the structure and organization of a local avian haemosporidian assemblage (genera Plasmodium and Haemoproteus) in the Cerrado biome of Central Brazil for 5 years. We obtained 790 blood samples from 54 bird species of which 166 (21%) were infected with haemosporidians based on molecular diagnostics. Partial sequences of the parasite cytochrome b gene revealed 18 differentiated avian haemosporidian lineages. We also analysed the relationship of life-history traits (i.e., nesting height, migration status, nest type, sociality, body mass, and embryo development period) of the 14 most abundant bird species with the prevalence of avian haemosporidia. It was found that host species that bred socially presented a higher prevalence of Haemoproteus (Parahaemoproteus) than bird species that bred in pairs. Thus, aspects of host behaviour could be responsible for differential exposure to vectors. The assemblage of avian haemosporidia studied here also confirms a pattern that is emerging in recent studies using molecular markers to identify avian haemosporidians, namely that many lineages are host generalists.

In stark contrast to the huge body of theoretical work on the importance of hosts and parasites as selective agents acting on each other, until recently, little systematic empirical investigation of this issue has been attempted. Research on snail–schistosome interactions have, therefore, the potential for making an important contribution to the study of coevolution or reciprocal adaptation. This may be particularly pertinent since snail–schistosomes represent an indirectly transmitted macroparasite system, so often overlooked amongst both theoretical and empirical studies. Here we review ideas and experiments on snail–schistosome interactions, with particular emphasis on those that may have relevance to the potential coevolution between host resistance and parasite infectivity and virulence. We commence with an introduction and definition of the general concepts, before going into detail of some specific studies to illustrate these: evidence of snail–schistosome coevolutionary process in the field; evidence of coevolutionary processes in the laboratory; a general assessment of the applicability of coevolutionary models in snail–schistosome interactions; and finishing with a section on conclusions and areas for further study.

Leishmaniasis is a vector-borne infectious disease caused by multiple Leishmania (L.) species with diverse clinical manifestations. There is currently no vaccine against any form of the disease approved in humans, and chemotherapy is the sole approach for treatment. Unfortunately, treatment options are limited to a small number of drugs, partly due to high cost and significant adverse effects. The other obstacle in leishmaniasis treatment is the potential for drug resistance, which has been observed in multiple endemic countries. Immunotherapy maybe another important avenue for controlling leishmaniasis and could help patients control the disease. There are different approaches for immunotherapy in different infectious diseases, generally with low-cost, limited side-effects and no possibility to developing resistance. In this paper, different immunotherapy approaches as alternatives to routine drug treatment will be reviewed against leishmaniasis.

Little is known about the effects of infection with Borrelia burgdorferi, the bacterium that causes Lyme disease, on its tick vectors. The purpose of this study was to determine the behavioural and ecological effects of infection by the bacterium in nymphal and adult black-legged (Ixodes scapularis) ticks. We found that the effects of infection were more pronounced in adults than in nymphs. Compared to uninfected adults, infected adults were less able to overcome physical obstacles, avoided vertical surfaces, were less active and quested at lower heights. Infected nymphs showed increased phototaxis and attraction to vertical surfaces. Infected nymphs also showed trends toward increased questing height and a greater tendency to overcome physical obstacles although these trends were not statistically significant. These altered behaviours in an infected tick may affect survival or pathogen transmission and may reflect kin selection in the bacterial pathogen.

A mathematical model of the population biology of Ostertagia ostertagi is described. The model assumes that the natural control and regulation of parasite numbers is mediated by four processes: the effect of climate on the development and survival of the free-living stages; changes in the rate of establishment of the infective larvae, and density-dependent variations in parasite survival and fecundity. The model is used to compare the course of the infection in two groups of calves. One group is assumed to have been reared under conditions typical of S.E. England and the other under conditions typical of Louisiana, USA. The more general behaviour of the model is discussed in the context of sensitivity analysis. The model is an excellent mimic of the epidemiology of bovine ostertagiasis. It can be used as a simple screening procedure to help determine which of many possible anthelmintic control strategies should be selected for more detailed examination in the field, and it provides a theoretical framework within which ideas concerning the epidemiology of parasitic gastro-enteritis can be assessed and refined.

Ascaris lumbricoides, the human roundworm, is a remarkably infectious and persistent parasite. It is a member of the soil-transmitted helminths or geohelminths and infects in the order of 1472 million people worldwide. Despite, its high prevalence and wide distribution it remains along with its geohelminth counterparts, a neglected disease. Ascariasis is associated with both chronic and acute morbidity, particularly in growing children, and the level of morbidity assessed as disability-adjusted life years is about 10·5 million. Like other macroparasite infections, the frequency distribution of A. lumbricoides is aggregated or overdispersed with most hosts harbouring few or no worms and a small proportion harbouring very heavy infections. Furthermore, after chemotherapeutic treatment, individuals demonstrate consistency in the pattern of re-infection with ascariasis, described as predisposition. These epidemiological phenomena have been identified, in a consistent manner, from a range of geographical locations in both children and adults. However, what has proved to be much more refractory to investigation has been the mechanisms that contribute to the observed epidemiological patterns. Parallel observations utilizing human subjects and appropriate animal model systems are essential to our understanding of the mechanisms underlying susceptibility/resistance to ascariasis. Furthermore, these patterns of Ascaris intensity and re-infection have broader implications with respect to helminth control and interactions with other important bystander infections.

Within the World Health Organization 2012–2020 roadmap for control and elimination of schistosomiasis, the scale-up of mass drug administration with praziquantel is set to change the epidemiological landscape across Africa and Arabia. Central in measuring progress is renewed emphasis upon diagnostics which operate at individual, community and environmental levels by assessing reductions in disease, infections and parasite transmission. However, a fundamental tension is revealed between levels for present diagnostic tools, and methods applied in control settings are not necessarily adequate for application in elimination scenarios. Indeed navigating the transition from control to elimination needs careful consideration and planning. In the present context of control, we review current options for diagnosis of schistosomiasis at different levels, highlighting several strengths and weaknesses therein. Future challenges in elimination are raised and we propose that more cost-effective diagnostics and clinical staging algorithms are needed. Using the Kingdom of Saudi Arabia as a contemporary example, embedding new diagnostic methods within the primary care health system is discussed with reference to both urogenital and intestinal schistosomiasis.