1. The phosphatases present in the adult and cysticercus stages of Taenia pisiformis have been investigated using histochemical and biochemical methods.

2. Histochemical tests failed to demonstrate the sites of enzyme activity in the cysticercus.

3. In the adult, the acid phosphatase is confined to the cuticle. Alkaline phosphatase occurs in the cuticle, subcuticular cells and the membranes bounding the ovary and vitelline tubules.

4. The histochemical distribution is uneven along the length of the worm, both acid and alkaline phosphatase being predominant hi the region of ‘mature’ proglottides. The scolex was negative to both tests.

5. Biochemical tests have demonstrated distinct acid and alkaline phosphatases in the cysticercus and adult stages. In the cysticercus the acid enzyme is predominant and in the adult it is the alkaline, implying a change in relative magnitude during the completion of the life cycle.

6. pH-activity curves have been obtained for the enzymes of both stages.

7. The results are discussed in relation to recent findings in the field of cestode enzymology, and it is suggested that these phosphatases may be associated with active transport of materials across the cuticle and ovarian and vitelline membranes.

Three populations of small wild British rodents were studied by capture–recapture methods over a period of 3 years, a fourth group was studied for 1 year and a fifth was sampled annually for 4 years. Blood smears were taken from 3 species of rodents: the woodmouse A podemus sylvaticus, the bank vole Clethrionomys glareolus (and an island sub-species C.g. skomerensis) and the short-tailed vole Microtus agrestis. The smears were examined microscopically. Four genera of haemoparasites Babesia, Hepatozoon, Trypanosoma and Grahamella were detected. Babesia was absent from C.g. skomerensis, Hepatozoon was rarely found in A. sylvaticus and M. agrestis and Trypanosoma was rare in A. sylvaticus. More males were infected than females but the difference was only statistically significant for the infection with Hepatozoon in adult C.g. skomerensis. Infections with Babesia and Hepatozoon were more prevalent in adult animals and infections with Trypanosoma were more prevalent in younger individuals. Only in C.g. skomerensis was there a significant difference between age classes in the prevalence of infection with Grahamella – there being more adults infected. Concurrent infections were detected, Hepatozoon being the parasite most commonly involved. The prevalence of infections was found to be approximately proportional to the number of animals known to be alive, regardless of the season.

Vertically transmitted parasites are transmitted from generation to generation of hosts usually via the host's gametes. Owing to gamete size dimorphism, the major transmission route is transovarial and selection (on the parasite) favours strategies which increase the relative frequency of the transmitting (female) host sex. These strategies impose unusual selection pressures on the host, and coevolution between hosts and vertically transmitted parasites has been implicated in speciation, in the evolution of symbiosis, and in the evolution of novel systems of host reproduction and sex determination. We review the evolutionary implications of vertically transmitted parasites in arthropods before focusing on strategies of transmission of a parasitic sex ratio distorter in Gammarus duebeni. The efficiency of parasite transmission to new hosts is a key factor underlying the relationship between vertically transmitted parasites and their hosts. Vertically transmitted parasites must overcome 2 bottlenecks in order to ensure successful infection of future host generations: first, transmission from adult to gamete; and secondly, transmission to the germ-line of the infected host. We investigate these 2 processes with regard to transovarial transmission by a microsporidian parasite in Gammarus duebeni. Parasite transmission from adult to eggs is highly efficient, with 96% of eggs of infected mothers inheriting the infection, whereas transmission to germ-line within infected embryos is relatively inefficient (72%). We measure parasite distribution between cells of developing embryos, and use these distributions to infer possible mechanisms of parasite transmission to germ-line. Parasite distribution within the embryo is dependent on host cell lineage, and is not consistent with unbiased segregation between daughter cells. These results indicate that parasites segregate together at host cell division, and may reflect a strategy of differential segregation to the host germ-line. We consider alternative parasite strategies at the cell-level in terms of their evolutionary implications.

The life cycle of Coitocaecum anaspidis involves three important stages. The adult occurs in Galaxias brevipennis, G. attentuatus, Gobiomorphus gobioides, and in specimens of Anguilla spp. less than 30 cm. long. Potamopyrgus antipodum and P. badia are the molluscan hosts of the sporocyst and cercaria. In New Zealand the cercaria attacks and encysts in Paracalliope fluviatilis, and in Australia in Anaspides tasmaniae. A metamorphosis of the cercarial larva into a progenetic metacercaria takes place in the amphipod.

Paracalliope does not become infected with Coitocaecum till the crustacean has reached 2·0 mm. in length. The thigmotropism of the Paracalliope fits exactly with the ground-living habit of the stumpy-tailed cercaria. The small river fish feeding in slack water acquire Coitocaecum from Paracalliope.

The risk factors associated with gametocytaemia at presentation and after treatment with different antimalarial drug regimens were evaluated in 767 children enrolled prospectively in 5 antimalarial drug trials between July 1996 and December 2002 in a hyperendemic area of southwestern Nigeria. The children were assigned to one of 6 treatment groups: chloroquine (CQ) only; pyrimethamine-sulfadoxine (PS) only; amodiaquine (AQ) only; CQ combined with chlorpheniramine (CQCP); or PS combined with CQ (CQPS) or AQ (AQPS). At enrolment, 115 (15%) of 767 children were gametocyte carriers. During follow-up, 15·6% of all patients (i.e. 120 patients) developed patent gametocytaemia, which in 85% (102 patients) had developed by day 7 following treatment. In a multiple regression model, 4 factors were found to be independent risk factors for the presence of gametocytaemia at enrolment: male gender (adjusted odds ratio [AOR]=0·55, 95% confidence interval [CI] 0·36–0·83, P=0·005), absence of fever (AOR=1·61, 95% CI 1·05–2·5, P=0·03), duration of illness >3 days (AOR=1·57, 95% CI 1·0–2·4, P=0·047), and asexual parasite densities less than 5000/μl (AOR=0·42, 95% CI 0·24–0·73, P=0·002). The presence of patent gametocytaemia at enrolment (AOR=0·04, 95% CI 0·02–0·07, P<0·001) and recrudescence of asexual parasites within 14 days were associated with the presence of gametocytaemia 7 or 14 days after enrolment (AOR=0·5, 95% CI 0·3–0·8, P=0·007). Delay in the time taken to clear the initial parasitaemia (>2 days) was associated with increased risk of subsequent gametocyte carriage. These findings may have implications for malaria control efforts in sub-Saharan Africa where control of the disease depends almost entirely on chemotherapy.

We have used immunoelectron microscopy to investigate the subcellular location of Pfs16 in Plasmodium falciparum. It was detected in the outer membrane region of gametocytes and more specifically on the parasitophorous vacuole membrane (pvm), since, during gametogenesis when the pvm disintegrates, the majority of the antigen was detected on the remains of this membrane in multilaminated whorls and not on the gamete plasma membrane. The antigen was also present on other gametocyte cellular structures, including those which we believe to be Garnham bodies, present in the host cell cytoplasm of some gametocytes. The antigen was present too on the membrane surrounding cytostomes and the resulting food vacuoles in the parasite cytoplasm.

Juvenile hedgehogs having insufficient body weight are often brought for overwintering to hedgehog rehabilitation centres. Faecal samples of juvenile hedgehogs and overwintering hedgehogs (n=188) collected prior to releasing them back into the wilderness were examined for the presence of Cryptosporidium coproantigen and oocysts. Altogether 56 (29·8%) submitted samples were positive for coproantigen. Forty-five (39·5%, n=114) of the positive samples originated from newly rescued hedgehogs, while 11 (14·8%, n=74) positive samples were from animals that spent several months at the station. Fifteen samples subjected to PCR-RFLP analysis on the partial 18S rRNA locus suggested the presence of C. parvum. Multilocus sequence typing on partial 60 kDa glycoprotein gene, 18S rRNA, actin gene, 70 kDa heat shock protein gene sequences revealed 3 different subtype families: IIa, IIc and a new, proposed as VIIa subtype family. Cryptosporidium sp. genotype belonging to VIIa subtype family is closely related to C. parvum but is genetically distinct being probably a hedgehog-specific Cryptosporidium sp. genotype with unknown zoonotical potential. Hedgehogs excreting Cryptosporidium oocysts represent a potential source for human infections, but also an anthroponotic nature of the IIc subtype family should be reviewed.

Population genetics of multi-host pathogens offers great potential for the understanding of their complex epidemiology but care must be taken to ensure that the sampling procedure does not bias estimates of population indices. The transfer of material to laboratory passage, in particular, runs the risk of bottlenecking and imposing non-random host-induced selection pressures according to the hosts used in passage. We present a novel technique allowing single-locus microsatellite genotyping of the naturally sampled larval stages, enabling unbiased population genetic studies of the multi-host zoonotic parasite Schistosoma japonicum. The utility of these larval genotyping methods for molecular epidemiological studies are illustrated in results from 3 separate data sets. In the first data set, potential loss of alleles based on the definitive host species used for laboratory maintenance was identified by comparing adult worm populations derived from mice and rabbits infected with cercarial populations originating from the same set of snails. In the second data set, bottlenecking was demonstrated by the loss of alleles in adult worms derived within a single generation of laboratory maintenance compared to their parent field-collected cercarial samples. In the final data set, comparison of miracidia and adult worms recovered from naturally infected animals demonstrated that larval analyses can provide stage-specific epidemiological information and that population genetics of schistosomes can be well described by analysis of larval stages. Our results thus advocate the use of natural life-cycle stages to obtain an accurate and ethical representation of the population genetic structure of S. japonicum and other multi-host pathogens.

The zoonotic cestode Echinococcus ortleppi (Lopez-Neyra and Soler Planas, 1943) is mainly transmitted between dogs and cattle. It occurs worldwide but is only found sporadically in most regions, with the notable exception of parts of southern Africa and South America. Its epidemiology is little understood and the extent of intraspecific variability is unknown. We have analysed in the present study the genetic diversity among 178 E. ortleppi isolates from sub-Saharan Africa, Europe and South America using the complete mitochondrial cox1 (1608 bp) and nad1 (894 bp) DNA sequences. Genetic polymorphism within the loci revealed 15 cox1 and six nad1 haplotypes, respectively, and 20 haplotypes of the concatenated genes. Presence of most haplotypes was correlated to geographical regions, and only one haplotype had a wider spread in both eastern and southern Africa. Intraspecific microvariance was low in comparison with Echinococcus granulosus sensu stricto, despite the wide geographic range of examined isolates. In addition, the various sub-populations showed only subtle deviation from neutrality and were mostly genetically differentiated. This is the first insight into the population genetics of the enigmatic cattle adapted Echinococcus ortleppi. It, therefore, provides baseline data for biogeographical comparison among E. ortleppi endemic regions and for tracing its translocation paths.

Metacyclic forms of Leishmania major and putative metacyclics of L. mexicana mexicana were found to occur in abundance in stationary phase cultures. These forms have been compared in several ways with promastigotes from mid-log phase cultures and, in the case of L. m. mexicana, amastigotes. Metacyclics are smaller, contain less protein and appear more active than other promastigotes. Both forms of promastigote respire at a high rate in the absence of exogenous substrate. The free amino-acid contents of the various forms of the two species have been analysed. They differ in detail but alanine was the major amino acid in all cases. The isoenzyme content of the different forms differed significantly. That of the putative metacyclics of L. m. mexicana was in several respects more similar to amastigotes than promastigotes, suggesting that the form is pre-adapted for life in a mammal. Metacyclics of L. major apparently did not divide in culture but transformed back over a period of 48 h to mid-log phase cells. The results provide further detail of the molecular differences between mid-log phase and metacyclic promastigotes and confirm that metacyclics are a distinct form in the life-cycle.

This paper summarises the progress towards vaccine development against the major blood-feeding nematodes of man and livestock, the hookworms and Haemonchus contortus, respectively. The impact of the diseases and the drivers for vaccine development are summarized as well as the anticipated impact of the host immune response on vaccine design. The performance requirements are discussed and progress towards these objectives using defined larval and adult antigens, many of these being shared between species. Specific examples include the Ancylostoma secreted proteins and homologues in Haemonchus as well as proteases used for digestion of the blood meal. This discussion shows that many of the major vaccine candidates are shared between these blood-feeding species, not only those from the blood-feeding stages but also those expressed by infective L3s in the early stages of infection. Challenges for the future include: exploiting the expanding genome information for antigen discovery, use of different recombinant protein expression systems, formulation with new adjuvants, and novel methods of field testing vaccine efficacy.

A study carried out on a sheep farm examined whether Toxoplasma gondii foetal infection and associated abortion occur in successive lambings. We identified 29 ewes that gave birth to lambs on at least 2 successive years over our study period, 2000–2003. Tissue samples from the progeny of these ewes were analysed by PCR to determine infection status with T. gondii. T. gondii-infected lambs were born in 31% of successive pregnancies. T. gondii-positive lambs were aborted in successive pregnancies in 21% of lambings during study period, 2000–2003. The frequency of successive abortions within this flock over the period 1992–2003 was 18%. If a lamb was congenitally infected there was a high risk (69%) that the successive lamb from that ewe would also be congenitally infected. Similarly, if a lamb was aborted there was a high risk (55%) of abortion in the next lamb produced. These data suggest that life-long immunity to T. gondii infections may not always be acquired following an initial infection and raises the question as to whether the mechanisms of T. gondii transmission prior to and during ovine pregnancies are fully understood.

Studies incorporating the ecology of clinical and sub-clinical disease in wild populations of conservation concern are rare. Here we examine sub-clinical infection by Trichomonas gallinae in a declining population of free-living European Turtle Doves and suggest caseous lesions cause mortality in adults and nestlings through subsequent starvation and/or suffocation. We found a 100% infection rate by T. gallinae in adult and nestling Turtle Doves (n = 25) and observed clinical signs in three adults and four nestlings (28%). Adults with clinical signs displayed no differences in any skeletal measures of size but had a mean 3·7% reduction in wing length, with no overlap compared to those without clinical signs. We also identified T. gallinae as the suggested cause of mortality in one Red-legged Partridge although disease presentation was different. A minimum of four strains of T. gallinae, characterized at the ITS/5·8S/ITS2 ribosomal region, were isolated from Turtle Doves. However, all birds with clinical signs (Turtle Doves and the Red-legged Partridge) carried a single strain of T. gallinae, suggesting that parasite spill over between Columbidae and Galliformes is a possibility that should be further investigated. Overall, we highlight the importance of monitoring populations for sub-clinical infection rather than just clinical disease.

Ascaris suum contains a large number of FMRFamide-related peptides (FaRPs) of which KNEFIRFamide (AF1), KHEYLRFamide (AF2) and KSAYMRFamide (AF8, also called PF3) have been extensively studied and are known to exert actions on somatic muscle strips of the worm. In the present study, the effects of AF1, AF2 and AF8 on the activity of the vagina vera of female A. suum have been examined in vitro. The vagina vera is a muscular tube connecting the uterus and vagina uteri to the gonopore and is probably involved in regulating egg output. The tissue exhibited spontaneous, rhythmic contractions in vitro, which were modulated by each of the FaRPs tested. The effects of each of the peptides were qualitatively and quantitatively different, and in each case were reversible. AF1 (1 μM) caused a biphasic response in the form of a transient lengthening of the preparation, followed by a shortening; contractions were initially inhibited but resumed 5 min post-addition of the peptide. Lower concentrations ([les ]0·1 μM) induced a less marked effect, with rhythmic contractions returning 5 min post-addition. AF2 and AF8 reduced contraction frequency at concentrations [ges ]0·1 μM. Both peptides also caused the tissue to shorten, although the effects of AF8 on baseline tension were inconsistent. The apparent potencies of AF1 and AF8 on contraction frequency of the vagina vera were 10-fold greater than AF2 and, unlike their actions on A. suum somatic body wall muscles, the actions of AF1 and AF2 were qualitatively different. Indeed, the effects of each of these FaRPs on the vagina vera were markedly different from those observed on the somatic muscle.

Chagas disease is caused by infection with the insect-transmitted protozoan Trypanosoma cruzi, and is the most important parasitic infection in Latin America. The current drugs, benznidazole and nifurtimox, are characterized by limited efficacy and toxic side-effects, and treatment failures are frequently observed. The urgent need for new therapeutic approaches is being met by a combined effort from the academic and commercial sectors, together with major input from not-for-profit drug development consortia. With the disappointing outcomes of recent clinical trials against chronic Chagas disease, it has become clear that an incomplete understanding of parasite biology and disease pathogenesis is impacting negatively on the development of more effective drugs. In addition, technical issues, including difficulties in establishing parasitological cure in both human patients and animal models, have greatly complicated the assessment of drug efficacy. Here, we outline the major questions that need to be addressed and discuss technical innovations that can be exploited to accelerate the drug development pipeline.

Parasitic helminths belong to 3 separate phyla and there is always the danger of over-generalization. The various routes of anaerobic carbohydrate breakdown in parasitic helminth differ in their efficiencies and in their power output. The choice of end-product represents a compromise between these two conflicting forces. In addition, anaerobic pathways must satisfy the redox requirements of the tissues and provide a source of intermediates for synthetic reactions. Other considerations include the metabolic cost of excretion and the effect of end-products on protein structure and function. The different end-products may fulfil additional functions such as pH control, nitrogenous excretion, osmotic regulation, intracellular signalling and the suppression of host responses.

A complicating factor in parasitic helminths is the existence of strains with different biochemical characteristics, including marked variation in end-product formation. The various tissues of the same parasite can also produce different end-products and the pattern of end-product formation is influenced by a variety of extrinsic and intrinsic factors such as age, sex, length of incubation, pO2 and availability of substrates. The catabolic pathways of helminths thus show considerable functional adaptation.

There is, as yet, no satisfactory explanation as to why helminths do not make the maximum use of any oxygen available to them; and the contribution of oxidative processes to the overall energy balance of parasites probably varies from species to species.

The catabolic pathways of adult helminths are derived from the anaerobic pathways present in their free-living relatives. Two main trends are evident, homolactic fermentation and carbon dioxide fixation, the latter involving a partial reverse tricarboxylic acid cycle. In general, homolactic fermentation is found in blood and tissue parasites, carbon dioxide fixation in gut parasites. These two types of metabolism are, of course, in no way absolute, most homolactic fermentors fix carbon dioxide to a certain extent and many parasites which fix carbon dioxide also produce lactate. Parasitic helminths possess a wide range of different catabolic pathways, superimposed upon which is a high degree of functional plasticity.

As training in helminthology has declined in the medical microbiology curriculum, many rare species of zoonotic cestodes have fallen into obscurity. Even among specialist practitioners, knowledge of human intestinal cestode infections is often limited to three genera, Taenia, Hymenolepis and Dibothriocephalus. However, five genera of uncommonly encountered zoonotic Cyclophyllidea (Bertiella, Dipylidium, Raillietina, Inermicapsifer and Mesocestoides) may also cause patent intestinal infections in humans worldwide. Due to the limited availability of summarized and taxonomically accurate data, such cases may present a diagnostic dilemma to clinicians and laboratories alike. In this review, historical literature on these cestodes is synthesized and knowledge gaps are highlighted. Clinically relevant taxonomy, nomenclature, life cycles, morphology of human-infecting species are discussed and clarified, along with the clinical presentation, diagnostic features and molecular advances, where available. Due to the limited awareness of these agents and identifying features, it is difficult to assess the true incidence of these ‘forgotten’ cestodiases as clinical misidentifications are likely to occur. Also, the taxonomic status of many of the human-infecting species of these tapeworms is unclear, hampering accurate species identification. Further studies combining molecular data and morphological observations are necessary to resolve these long-standing taxonomic issues and to elucidate other unknown aspects of transmission and ecology.

In 2009, a haplotype closely related to European strains of Echinococcus multilocularis was detected in a dog from the Quesnel region of British Columbia, Canada. We now report the establishment of this same haplotype in 7 coyotes (Canis latrans) trapped within 40 km of Quesnel, BC. In addition, 3 coyotes and 1 red fox (Vulpes vulpes) harboured adult cestodes morphologically compatible with that of E. multilocularis (overall prevalence 33% in 33 carnivores). None of 156 potential intermediate hosts, including 131 representatives of two highly suitable rodent species, Peromyscus maniculatus and Microtus pennsylvanicus, trapped from a region 120–210 km south of Quesnel were infected. This report confirms the establishment and local transmission of a European-type strain of E. multilocularis (the causative agent of human alveolar hydatid disease), in wildlife in a forested region of North America where this cestode had not been previously detected, with significance for public and animal health.

The life-cycles of two closely related cephalobaenid pentastomids, Raillietiella gehyrae and Raillietiella frenatus, which utilize geckos as definitive hosts and cockroaches as intermediate hosts, have been investigated in detail. Early development in the fat-body of cockroaches involves 2 moults to an infective, 3rd-stage larva which appears from 42–44 days post-infection. Complete development in geckos involves a further 5 moults in the case of males and 6 for females. Males mature precociously and copulation is a once-in-a-lifetime event which occurs around day 80 post-infection when both sexes are the same size but the uterus of the female is undeveloped. Sperm, stored in the spermathecae, is used to fertilize oocytes which slowly accumulate in the developing saccate uterus. Patency commences when the uterus carries approximately 4000–5500 eggs but only 25–36 % of these contain fully developed primary larvae. Since only mature eggs are deposited, we postulate that the vagina (?) of the female must be equipped with a selective filter that allows through large eggs but retains smaller, immature eggs. Thus the only limit on fecundity is the total number of sperms in the spermathecae and this is precisely the same factor that constrains egg production in the advanced order Porocephalida.

As a consequence of comprehensive transcriptome analysis followed by sequencing and draft assembly of the genome, the emphasis of schistosome research is shifting from the identification of genes to the characterization of their functions and interactions. Developmental biologists have long used whole mount in situ hybridization (WISH) to determine gene expression patterns, as a vital tool for formulating and testing hypotheses about function. This paper describes the application of WISH to the study of gene expression in larval and adult schistosomes. Fixed worms were permeablized by proteinase K treatment for hybridization with digoxygenin-labelled RNA probes, with binding being detected by alkaline phosphatase-coupled anti-digoxygenin antibodies, and BM Purple substrate. Discrete staining patterns for the transcripts of the molecules Sm29, cathepsin L, antigen 10.3 and chorion were observed in the tegument cell bodies, gut epithelium, oesophageal gland and vitelline lobules, respectively, of adult worms. Transcripts of the molecules SGTP4, GP18-22 and cathepsin L were localized to tegument cell bodies and embryonic gut, respectively, of lung schistosomula. We also showed that Fast Red TR fluorescent substrate can refine the pattern of localization permitting use of confocal microscopy. We believe that method of WISH will find broad application, in synergy with other emerging post-genomic techniques, such as RNA interference, to studies focused at increasing our molecular understanding of schistosomes.