Genomes of the major human helminth parasites, and indeed many others of agricultural significance, are now the research focus of intensive genome sequencing and annotation. A draft genome sequence of the filarial parasite Brugia malayi was reported in 2007 and draft genomes of two of the human schistosomes, Schistosoma japonicum and S. mansoni reported in 2009. These genome data provide the basis for a comprehensive understanding of the molecular mechanisms involved in schistosome nutrition and metabolism, host-dependent development and maturation, immune evasion and invertebrate evolution. In addition, new potential vaccine candidates and drug targets will likely be predicted. However, testing these predictions is often not straightforward with schistosomes because of the difficulty and expense in maintenance of the developmental cycle. To facilitate this goal, several developmental stages can be maintained in vitro for shorter or longer intervals of time, and these are amenable to manipulation. Our research interests focus on experimental studies of schistosome gene functions, and more recently have focused on development of transgenesis and RNA interference with the longer term aim of heritable gene manipulation. Here we review methods to isolate and culture developmental stages of Schistosoma mansoni, including eggs, sporocysts, schistosomules and adults, in particular as these procedures relate to approaches for gene manipulation. We also discuss recent advances in genetic manipulation of schistosomes including the deployment of square wave electroporation to introduce reporter genes into cultured schistosomes.

Within recent years, problems of host-parasite relationships have been receiving increasing attention from parasitologists. This is particularly true in the study of the cestodes Echinococcus spp., where predilection for different hosts—when considered with the morphology of the adult and the growth and development of the larva—seemed sufficiently significant to lead to the establishment of the two species, E. granulosus and E. multilocularis. Much experimental and field work has been carried out on a variety of vertebrate hosts and a number of new, naturally infected and laboratory hosts have been reported. A survey of intermediate and definitive hosts has been undertaken in this laboratory as part of a study to investigate whether factors of morphological, physiological or biochemical origin were concerned in host specificity. These data, together with some genetical considerations, form the basis of the present paper.

The effect of treatment with monoclonal antibodies (Mabs) which deplete CD4+ or CD8+ T lymphocytes, on infections with Eimeria spp. was examined in NIH mice. Treatment with anti-CD4 Mab increased susceptibility to primary infections with E. vermiformis or E. pragensis and reduced the subsequent resistance of the mice to homologous challenge. Similar treatment of immune mice did not affect their resistance to re-infection but this was reduced in mice depleted of CD8+ T lymphocytes. In mice immunized with E. vermiformis the effect of CD8+-depletion was very slight, apparent only as the presence of small numbers of oocysts in the faeces of some mice; in mice immunized with E. pragensis there was a small, though significant, increase in oocyst production, compared with controls and anti-CD4-treated groups. These results confirm the importance of mechanisms involving the function of CD4+ T lymphocytes in the control of primary infections with Eimeria spp. and indicate that CD8+ cells play some part in the expression of resistance to reinfection. They also show that a major part of this resistance was not affected by either of the treatments given.

The ascaridoid nematode of cats from Kuala Lumpur, Malaysia, previously identified morphologically as Toxocara canis, was characterized using a molecular approach. The nuclear ribosomal DNA (rDNA) region spanning the first internal transcribed spacer (ITS-1), the 5·8S gene and the second internal transcribed spacer (ITS-2) was amplified and sequenced. The sequences for the parasite from Malaysian cats were compared with those for T. canis and T. cati. The sequence data showed that this taxon was genetically more similar to T. cati than to T. canis in the ITS-1, 5·8S and ITS-2. Differences in the ITS-1 and ITS-2 sequences between the taxa (9·4–26·1%) were markedly higher than variation between samples within T. canis and T. cati (0–2·9%). The sequence data demonstrate that the parasite from Malaysian cats is neither T. canis nor T. cati and indicate that it is a distinct species. Based on these data, PCR-linked restriction fragment length polymorphism (RFLP) and single-strand conformation polymorphism (SSCP) methods were employed for the unequivocal differentiation of the Toxocara variant from T. canis and T. cati. These methods should provide valuable tools for studying the life-cycle, transmission pattern(s) and zoonotic potential of this parasite.

The morbidity and transmission dynamics of geohelminthiases are determined by the patterns of infection intensity in the community. Understanding the determinants of these patterns requires a combination of field, laboratory and theoretical study. Studies of age-specific reinfection, and of the phenomenon of predisposition, indicate that the major determinant of convex age-intensity profiles and of heterogeneity in infection intensity is the rate of establishment of infection, rather than the rate of adult worm mortality. The rate of establishment is, in turn, determined by exposure to, and protection from, infection. The evidence indicates that exposure, at least to the orally-transmitted geohelminths, varies with age and is highly heterogeneous between hosts. The immune response in geohelminthiasis is vigorous, parasite-specific, hetero geneous between hosts, and both age and infection dose dependent, but has yet to be convincingly shown to be protective. Since the immune response is itself a function of exposure, unravelling the interaction between ecology and immunology as determinants of geohelminth worm burden will require simultaneous assessment of both processes via immuno epidemiological study.

Following a period characterized by severe epidemics of sleeping sickness, restoration of effective control and surveillance systems has raised the question of eliminating the disease from sub-Saharan Africa. Given sufficient political and financial support, elimination is now considered a reasonable aim in countries reporting zero or less than 100 cases per year. This success may lead health authorities across the affected region to downgrade the disease from ‘neglected’ to simply being ignored. In view of the significant levels of under-reporting of sleeping sickness mortality in rural communities, this could be a short-sighted policy. Loss of capacity to deal with new epidemics, which can arise as a consequence of loss of commitment or civil upheaval, would have serious consequences. The present period should be seen as a clear opportunity for public-private partnerships to develop simpler and more cost-effective tools and strategies for sustainable sleeping sickness control and surveillance, including diagnostics, treatment and vector control.

Among the topics of considerable interest concerning our understanding of the unusual biology of schistosomes is the sexual maturation of the female. The identification of genes coding for signal transduction proteins controlling essential steps of the pairing-dependent differentiation of the reproductive organs, vitellarium and ovary will help to substantiate our knowledge about this unique parasite. Furthermore, such signalling proteins could be potential targets to interfere with the development of this parasite to combat schistosomiasis since its pathology is caused by the eggs. This review summarises first post-genomic steps to elucidate the function of gonad-specific signalling molecules which were identified by homology-based cloning strategies, by in silico identification or by yeast two-hybrid interaction analyses, using a combination of novel techniques. These include the in vitro culture of adult schistosomes, their treatment with chemical inhibitors to block enzyme activity, the use of RNAi to silence gene function post-transcriptionally, and confocal laser scanning microscopy to study the morphological consequences of these experimental approaches. Finally, we propose a first model of protein networks that are active in the ovary regulating mitogenic activity and differentiation. Some of these molecules are also active in the testes of males, probably fulfilling similar roles as in the ovary.

Parasitism is defined in various ways as an intimate relationship in which one partner, the parasite, lives on or in another, the host, generally at the expense of the latter. Parasitism commonly results in a unique array of host physiological responses and adaptations. Most studies of the physiological effects of parasitism have focused on the pathological consequence of infection and disease. While many physiological changes contribute to pathogenesis, it is now recognized that parasitic infections at sub-clinical levels also produce physiological effects that either ameliorate or may not contribute to the disease process. Moreover, these physiological changes are often manifested by altered host behaviour. Behavioural studies have enabled an ecological- and evolutionary-oriented evaluation of host responses. In this fashion, physiological effects may be assessed as to whether they affect fitness and confer benefit or harm to one or both of the symbionts involved. We briefly examine how these physiological responses, specifically neural, endocrine, neuromodulatory, and immunomodulatory components, may interact to modify host behaviours. We consider the adaptiveness of these responses and how the behavioural patterns elicited may simultaneously appear adaptive for the parasite as well as the host. In addition, we address how parasite-host physiological and behavioural interactions may be altered during the course of parasitism.

Proliferative kidney disease (PKD), a condition associated with high mortality in salmonid fish, represents an abnormal immune response to the presence of an enigmatic myxozoan, which has been designated simply as PKX organism because its generic and specific status are obscure. Phylogenetic analyses of partial sequences of the 18S rDNA of PKX and of myxozoan parasites infecting the bryozoans Cristatella mucedo, Pectinatella magnifica and Plumatella rugosa, including the previously named Tetracapsula bryozoides from C. mucedo, showed that these taxa represent a distinct clade that diverged early in the evolution of the Myxozoa before the radiation of the other known myxozoan genera. A common feature of the myxozoans in this clade may be the electron-dense sporoplasmosomes with a lucent bar-like structure, which occur in T. bryozoides and PKX but not in the myxozoans belonging to the established orders Bivalvulida and Multivalvulida. Variation of 0·5–1·1% was found among the PKX 18S rDNA sequences obtained from fish from North America and Europe. The 18S rDNA sequence for T. bryozoides showed that it is a distinct taxon, not closely related to PKX but some sequences from myxozoans infecting 2 of the bryozoan species were so similar to those of PKX as to be indistinguishable. Other sequences from the new myxozoans in bryozoans at first appeared distinct from PKX in a maximum likelihood tree but, when analysed further, were also found to be phylogenetically indistinguishable from PKX. We propose that at least some variants of these new myxozoans from bryozoans are able to infect and multiply in salmonid fish, in which they stimulate the immune reaction and cause PKD but are unable to form mature spores to complete their development.

Studies on the digenean parasites of deep-sea (> 200 m depth) teleosts are reviewed and two case study generic phylogenies are presented based on LSU rDNA and ND1 mtDNA sequences. The phylogeny of the lepocreadiid genus Lepidapedon, the most common deep-sea digenean genus, is not clearly resolved as the two gene trees are not compatible. It can be inferred, however, that the genus has radiated in the deeper waters off the continental shelf, mainly in fishes of the gadiform family Macrouridae. Steringophorus, a fellodistomid genus, is better resolved. In this case a deep-sea radiation is also indicated, but the pattern of host-specificity is not clear, with evidence of much host-switching. Results of studies of the parasites of the macrourid fish Coryphaenoides (Nematomirus) armatus from various depths have reinforced recent views on the lack of zoned depth-related communities in the deep-sea. The diversity of deep-sea digeneans is relatively low with only 18 families (of about 60) reported. Little, or nothing, is known from most deep-sea areas and nothing from trenches and mid-ocean ridge systems.

The taxonomy of Echinococcus has long been controversial. Based mainly on differences in morphology and host-parasite specificity characteristics, 16 species and 13 subspecies were originally described. Subsequently, most of these taxa were regarded as synonyms for Echinococcus granulosus and only 4 valid species were recognised: E. granulosus; E. multilocularis; E. oligarthrus and E. vogeli. But, over the past 50 years, laboratory and field observations have revealed considerable phenotypic variability between isolates of Echinococcus, particularly those of E. granulosus, which include differences in: morphology in both larval and adult stages, development in vitro and in vivo, host infectivity and specificity, chemical composition, metabolism, proteins and enzymes, pathogenicity and antigenicity. The application of molecular tools has revealed differences in nucleic acid sequences that reflect this phenotypic variation and the genetic and phenotypic characteristics complement the previous observations made by the descriptive parasitologists many years ago. The fact that some of these variants or strains are poorly or not infective to humans has resulted in a reappraisal of the public health significance of Echinococcus in areas where such variants occur. A revised taxonomy for species in the Echinococcus genus has been proposed that is generally accepted, and is based on the new molecular data and the biological and epidemiological characteristics of host-adapted species and strains.

Infection of intermediate hosts with eggs of taeniid parasites results in a larval infestation known as cysticercosis. A number of studies have indicated that cysticercosis is associated with immunosuppression, although little is known about the mechanisms involved. In the present study, mice infected with the larvae of Taenia crassiceps were found to exhibit a pronounced anergy, which preferentially affected T-cells located anatomically close to the parasite. This anergy was linked to late events in the T cell activation pathway; that is, stimulation through the T cell receptor(TCR)/CD complex by Concanavalin-A, or plate-bound monoclonal antibodies (mAb) to TCRαβ or CD3ε, or combinations of phorbol ester and ionomycin (all of which can bypass early membrane-related events), failed to fully activate T lymphocytes. The relative proximity of T cells to the parasite was directly related to upregulation of IL-4 and downregulation of IL-2 production. In addition, the profiles of parasite-specific Abs showed an exclusive increase of serum IgG1 during infection. Taken together, the data suggest that infection of mice with larvae of T. crassiceps alters the balance of CD4+ Th cells by upregulating Th2 and downregulating Th1 cells located in close proximity to the parasite.

The development of the feeding lesions of the ixodid tick Boophilus microplus was followed throughout the parasitic life-cycle on cattle of both Zebu and European breeds, using histological techniques.

The feeding lesion of this tick must be the result of an interaction between the tick and its host as it is neither the result of simple mechanical trauma, nor of lysis of the host tissues by salivary secretions of the tick alone, nor is it solely a result of an inflammatory response.

The histological evidence suggests that the diet of the tick in each instar changes as the lesion develops; first, tissue fluids are available, then tissue fluids along with leucocytes which have infiltrated the lesion, and finally these two components along with whole blood.

An early intense infiltration of eosinophils into the area of the mouthparts, typical of an immediate hypersensitive or anaphylactic response, was noted with European cattle on secondary and subsequent exposures to larvae. Hypersensitivity to tick infestation, which has been equated with resistance to infestation, was seen as an extreme of a normal spectrum of sensitization which results in an abnormal susceptibility.

The definitive lesion was similar with all hosts studied. It consisted of a cavity containing leucocytes, predominantly neutrophils, some lymphocytes and erythrocytes in an area of heavily infiltrated collagen.

It is concluded that this host-parasite relationship illustrates the concept of adaptation tolerance as proposed by Sprent (1962).

It is with much pleasure that we acknowledge the continued generous financial support by the Australian Meat Research Committee (formerly the Australian Cattle and Beef Research Committee), which has made possible this project.

The development of malaria due to Plasmodium falciparum is a complex, multi-stage process. It is usually characterized by an exponential growth in the number of parasite-infected erythrocytes, followed by marked oscillations in this number with a period of 48 h, which are eventually dampened. This course of events has been the subject of various mathematical models. In this paper we propose a new mathematical model for the in-host asexual erythrocytic development of P. falciparum malaria. Synchronicity of the infection is shown to be an inherent feature of infection, irrespective of the duration of merozoite release from the liver. It will, therefore, cause periodic symptoms, as known in malaria patients. We also simulate the effects of an induced host immune response and show how the level of immunity affects the development of disease. The simulations fit well with the clinical observations. We show how infection can become asynchronous and discuss the effect of desynchronization on the circulating and total parasitaemia and demonstrate that synchronized broods will show parasitaemia fluctuations.

The timing of the energetic consequences of a developing, single-dose infection of Trichostrongylus tenuis larvae was investigated in captive red grouse Lagopus lagopus scoticus. At 12 days post-infection (p.i.), infected birds had a resting metabolic rate 16% greater than controls and thenceforth lost weight at a faster rate than controls. At 16 days p.i. infected birds consumed 38% less energy and excreted 33% less energy than controls. The estimated total daily energy expenditure and energy expended on activity for infected birds at 16 days p.i. were 36% and 83% lower, respectively, than for controls. Infected birds lost condition from 16 days p.i. onwards. The period of energy imbalance at 12–16 days p.i. coincided with development of late 4th-stage larvae into adult worms and the onset of patency. After this, the effects on energy balance diminished. Synchronous development of previously arrested T. tenuis larvae in wild birds in spring probably has similar effects to those reported here and places grouse under conditions of energy imbalance. The observed effects on energy balance provide a possible mechanism by which the parasite can reduce fecundity and survival of infected grouse.

The localization and distribution of neuropeptides in the nervous system of the liver fluke, Fasciola hepatica at different stages in the development of the adult fluke have been determined by an indirect immunofluorescence technique, using antisera to 19 vertebrate peptides and the invertebrate neuropeptide, FMRFamide. Positive immunoreactivity was obtained with antisera to pancreatic polypeptide (PP), peptide tyrosine tyrosine (PYY), substance P (SP) and FMRFamide. Cell bodies and nerve fibres immunoreactive to the 4 peptides are present in the anterior ganglia and the 3 pairs of longitudinal nerve cords and their commissures in the central nervous system. In the peripheral nervous system, immunoreactivity occurs in the nerve plexuses supplying the subtegumental musculature, the oral and ventral suckers, and the muscular lining of the male and female reproductive ducts, including the ootype, uterus, cirrus pouch and gonopore. Cells displaying immunoreactivity to PYY and FMRFamide lie amongst the Mehlis' gland cells that surround the ootype. Processes from these cells extend into the wall of the ootype. One group of PP-immunoreactive cells occurs at the junction of the vitelline and ovovitelline ducts, whilst another group is situated at the entrance to the uterus from the ootype. The results are discussed in relation to the possible roles of the peptides in the neurophysiology and egg production of the fluke.

Genes encoding small subunit ribosomal RNA (SSUrRNA) of 16 Blastocystis isolates from humans and other animals were amplified by the polymerase chain reaction, and the corresponding fragments were cloned and sequenced. Alignment of these sequences with the previously reported ones indicated the presence of 7 different sequence patterns in the highly variable regions of the small subunit ribosomal RNA. Phylogenetic reconstruction analysis using Proteromonas lacertae as the outgroup clearly demonstrated that the 7 groups with the different sequence patterns are separated to form independent clades, 5 of which consisted of the Blastocystis isolates from both humans (B. hominis) and other animals. The presence of 3 higher order clades was also clearly supported in the phylogenetic tree. However, a relationship among the 4 groups including these 3 higher order clades was not settled with statistical confidence. The remarkable heterogeneity of small subunit ribosomal RNAs among different Blastocystis isolates found in this study confirmed, with sequence-based evidence, that these organisms are genetically highly divergent in spite of their morphological identity. The highly variable small subunit ribosomal RNA regions among the distinct groups will provide useful information for the development of group-specific diagnostic primers.

Over 90% of Hymenolepis diminuta cysts administered by stomach tube become established in CFLP, and Porton strains of mice. The rate of growth of single worm infections in terms of dry weight and length was determined. The rate of growth of H. diminuta in mice is slower than in rats even in worms only a few mm long. In most mice growth ceases abruptly on day 10±1 day when the worm is 4–30 cm long, destrobilation usually occurs leaving a scolex and neck often 0·5–2 mm in length. Destrobilated worms can survive for at least 47 days but little if any regrowth occurs unless the worms are surgically transplanted into rats or previously uninfected mice. Normal growth appears to be resumed within hours in both recipient hosts. The cause of destrobilation is probably not physiological; position of establishment and forward migration during growth of the worm in mice is similar to that in a rat. Destrobilation occurs well before the worm fills the small intestine lengthwise. Strong evidence of an immunological response being involved is given by the fate of a secondary infection given 22 and 28 days after the primary. Lower % recoveries were recorded in secondary infections killed 6–9 days later, and the worms were severely stunted. The various lines of evidence are discussed and it is concluded no reasonable doubt exists that H. diminuta is rejected by CFLP and Porton mice as a result of an immunological response.

Most studies on the effects of tanniferous plants on nematodes have examined forages but have neglected the woody plants. Therefore, in vitro effects of extracts from 3 woody plants (Rubus fructicosus, Quercus robur, Corylus avellana) have been tested on trichostrongyles and compared to sainfoin, a legume forage. Because some in vivo results indicated that the effects of tannins differed depending on the parasitic species and/or stages, the effects were measured on 3rd-stage larvae (L3) and adult worms of Teladorsagia circumcincta, Haemonchus contortus and Trichostrongylus colubriformis. The effects of plant extracts varied according to the plant sources, the parasite species and stages. For the woody plants, significant inhibitory effects were obtained on both stages of abomasal species. Results for T. colubriformis were more variable. Effects of sainfoin extracts were significant on T. colubriformis and H. contortus L3, and on abomasal adult worms. In order to assess the implications of tannins, polyethylene glycol (PEG), an inhibitor of tannins, was added to hazel tree, oak and sainfoin extracts. Without PEG, significant inhibitory effects on L3 and adult worms were confirmed. After addition of PEG, the larval migration and motility of adult worms were restored in most cases. These results confirm variations in effects depending on factors related to plants or parasites and suggest that tannins are partly responsible for the effects.

Avian Plasmodium and Haemoproteus parasites are easily detected by DNA analyses of infected samples but only correctly assigned to each genus by sequencing and use of a phylogenetic approach. Here, we present a restriction site to differentiate between both parasite genera avoiding the use of those analyses. Alignments of 820 sequences currently listed in GenBank encoding a particular cytochrome B region of avian Plasmodium and Haemoproteus show a shared restriction site for both genera using the endonuclease Hpy CH4III. An additional restriction site is present in Plasmodium sequences that would initially allow differentiation of both genera by differential migration of digested products on gels. Overall 9 out of 326 sequences containing both potential restriction sites do not fit to the general rule. We used this differentiation of parasite genera based on Hpy CH4III restriction sites to evaluate the efficacy of 2 sets of general primers in detecting mixed infections. To do so, we used samples from hosts infected by parasites of both genera. The use of general primers was only able to detect 25% or less of the mixed infections. Therefore, parasite DNA amplification using general primers to determine the species composition of haemosporidian infections in individual hosts is not recommended. Specific primers for each species and study area should be designed until a new method can efficiently discriminate both parasites.