The species of Anisakis constitute one of the most widespread groups of ascaridoid nematodes in the marine ecosystem. Three closely related taxa are recognised in the A. simplex (s. l.) complex, i.e. A. pegreffii, A. simplex (s. s.) and A. berlandi. They are distributed in populations of their intermediate/paratenic (fish and squids) and definitive (cetaceans) hosts. A panel of seven microsatellite loci (Anisl 05784, Anisl 08059, Anisl 00875, Anisl 07132, Anisl 00314, Anisl 10535 and Anisl 00185), were developed and validated on a total of N = 943 specimens of A. pegreffii and A. simplex (s. s.), collected in fish and cetacean hosts from allopatric areas within the range of distribution of these parasite species. In addition, the locus Anisl 7, previously detected in those Anisakis spp., was investigated. The parasites were first identified by sequence analysis of the EF1 α-1 nDNA. The panel of the microsatellites loci here developed have allowed to: (i) detect diagnostic microsatellite loci between the two species; (ii) identify specimens of the two species A. pegreffii, A. simplex (s. s.) in a multi-marker nuclear genotyping approach; (iii) discover two sex-linked loci in both Anisakis species and (iv) estimate levels of genetic differentiation at both the inter- and intra-specific level.

The incidence of fever among infants in the village of Idete in the Morogoro region of Tanzania was analysed in relation to densities of Plasmodium falciparum parasites in the peripheral blood. Parasite densities in both fever cases and in asymptomatic infants, were compared and a Bayesian non-parametric mixture decomposition algorithm was used to estimate the proportion of fevers attributable to malaria and hence the incidence of clinical malaria. Age group-specific densities of peripheral parasitaemia showed little seasonality, but the clinical malaria incidence showed a clear peak in the wet season in children aged less than 9 months. Estimates of the parasitaemia-specific incidence of clinical malaria were used to quantify apparent tolerance of parasites, and indicated that clinical episodes occurred on average at lower parasite densities during the wet season than in the dry season. These patterns could reflect differences in levels of anti-toxic immunity, but the nature of the seasonal differences supports the alternative explanation that the variation in apparent tolerance may be an effect of changes in the ratio of peripheral parasite densities to the sequestered mass.

Ciné film and direct observation showed that microphagous, plant and animal nematodes have the same basic wave pattern when moving. Their speed is linearly related to the product of their length and wave frequency during unrestricted movement in deep water. The speed, wave pattern and frequency of the plant nematode Ditylenchus dipsaci and the animal nematode Trichostrongylus colubriformis changed in a similar way in response to changes in water film thickness on the surface of agar of different concentrations. T. colubriformis like D. dipsaci were most mobile in a sand-water medium at a suction corresponding to the stage when most of the water has drained from the pores. The optimum temperature for activity of T. colubriformis and D. dipsaci was about 20° C. and both survived desiccation at 50% relative humidity and 24° C. for 32 days. T. colubriformis was attracted to and aggregated around grass roots (Phleum pratense). The results suggest that the free-living stages of microphagous, plant parasitic and animal parasitic nematodes move similarly and that nematodes can be divided arbitrarily into three groups: (1) those active enough to swim in deep water; (2) those able to swim in thick water films, escape from the soil and ascend plants above ground level; and (3) those too inactive to swim, the crawlers, which are largely confined to the soil. The habitat of free-living stages of nematodes, irrespective of their host or feeding habits, is probably related to their propulsive power.

We thank Dr D. Poynter for supplying the animal parasites used.

The phylogenetic relationships of the Acanthocephala are reviewed in the light of recent ultrastructural evidence. Comparisons of spermatozoan ultrastructure suggest that the Acanthocephala cannot be closely related to the Platyhelminthes and that the similarities which exist between cestodes and acanthocephalans are the result of convergence. Similarities in the spermatozoan ultrastructure of acanthocephalans, rotifers, nematomorphs and nematodes probably have a phylogenetic significance and suggest that the acanthocephalans have a close common ancestor with some at least of the Aschelminthes. The case for this affinity is strengthened by ultrastructural evidence concerning the body walls and muscles of acanthocephalans, nematodes and rotifers.

I am indebted to Professor D. R. Arthur and Professor F. R. Whatley at King's College, London, for the provision of facilities. The work reported in this paper was undertaken during the tenure of a Research Fellowship from the Nuffield Foundation.

Cystoisospora belli is a coccidian protozoan that can cause chronic diarrhoea, acalculous cholecystitis and cholangiopathy in AIDS patients. We applied molecular methods to identify Cystoisospora at species level in AIDS patients presenting with and without the presence of unizoites in lamina propria. Coprological and histological analyses were performed in stool and/or biopsy samples from 8 Cystoisospora-infected patients. DNA from the same samples was used to amplify 2 fragments of the SSU-rRNA gene and the ITS-1 region. Sequencing of the resulting amplicons identified C. belli infections in all cases, independent of the presence or absence of unizoite tissue cysts. Further work should be considered in order to find molecular targets related to strain variations in C. belli.

An account is given of a virus which is present in the roots of a number of different plants. Plants which have been shown, so far, to contain the infective principle in the roots are several members of the Solanaceae, tomatoes, tobacco, etc.; of the Primulaceae, Polyanthus sp. and Primula obconica; of the Geraniaceae, zonal Pelargonium; and of the Compositae, the aster. Such plants show no indication of infection with the exception of Nicotiana tabacum and N. glutinosa, which may develop natural disease symptoms. These symptoms are usually confined to the two lowermost leaves and develop most frequently under conditions of low temperature and low light intensity. The virus never becomes completely systemic in any host plant and is generally restricted to the inoculated leaves. Experiments are described which show the virus to be present in the roots of tobacco plants which had given a negative virus reaction a few weeks earlier. The virus is found in the roots of tobacco plants arising from seed which has been heated to a temperature of 75° C., the thermal death-point of the virus in expressed sap being 72° C. All attempts to demonstrate the presence of the virus in the seed have proved negative.

Although no systematic study has been made, the range of plants susceptible to local infection in the leaves, by artificial inoculation, seems wide. Tests have shown that about 25 species of plants belonging to 10 families respond to inoculation with the virus.

Three possible explanations of the origin and spread of the infection are discussed.

The relevance of spatial variation in the environment and host communities for parasite community composition is poorly documented, creating a need for additional case studies from which general principles can be developed. Avian malaria in southern African waterfowl has not previously been studied. As a first step towards documenting and understanding its biogeography, we used PCR and molecular sequencing techniques to analyse 454 blood samples from Afrotropical ducks from 5 different locations (spread around the subregion) for avian malaria. Fifty-five blood samples were positive for one or more genera of haematozoa. The regional infection rate across all sites and sampling periods was 12·1%. Nine individuals carried dual infections containing multiple haematozoa. Fifteen different cytochrome b haplotypes among 52 positives (3 samples failed to sequence) and 61 total sequences were found. Eleven haplotypes closely matched Plasmodium, whereas 4 were more similar to Haemoproteus. Five distinct haematozoan clades were identified. Haemoproteus parasites appeared to be more host-specific than Plasmodium, which occurred at every sampling location and in every host species examined. There were no significant differences in overall parasite prevalence attributable to either site or species, although Plasmodium and Haemoproteus occurrences differed by site-species combination and the borderline significance of our test for between-site variation (P < 0·06) implied that with a larger sample size, differences in parasite prevalence among locations might be detectable.

The questing height (i.e. ambush height) of ticks on a plant plays an important role in host selection. To test the hypothesis that the questing height of ticks in a locality had adapted to the body size of the host in that locality, we examined the questing height of nymphs of the ticks, Haemaphysalis longicornis and H. mageshimaensis, at 7 locations in Japan. Sika deer, Cervus nippon, is the primary host of these ticks and there is considerable geographical variation in the body size of sika deer. Multiple regression analysis revealed that the questing height in the field was influenced by the height of the plants and by the body size of deer at a location. However, the questing height of ticks at some locations may have been constrained by the height of the plants and might not be the same as their intrinsic questing height. When ticks were placed in vertical glass tubes in the laboratory, the questing height of ticks from a locality was correlated with the mean body size of deer at that locality. Therefore, the prominent cue determining the questing height of H. longicornis and H. mageshimaensis seems to be the body size of the host deer.

Amoebae of the genus Endolimax have been found naturally inhabiting the large intestine of 3 species of Macacus (M. sinicus [= radiatus], M. rhesus [= mulatto], M. nemestrinus).

In all these hosts the amoebae are morphologically and culturally identical, at all stages of development, and are indistinguishable by any structural or cultural character from Endolimax nana of Man—with which they have been carefully compared. The several forms have been studied in “pure” culture (i.e., free from all other protozoa, but accompanied by bacteria suitable for their growth).

The Endolimax of M. sinicus, isolated in “pure” culture, has been experimentally transmitted to an uninfected man by the natural route (cysts per os): and in this human subject it has produced an infection indistinguishable from a natural one with E. nana. The induced infection has now persisted unchanged for close on 8 years.

From the experimentally infected man the amoeba has been artificially transmitted to an uninfected M. rhesus, in which it gave rise to an infection—likewise indistinguishable from a natural simian infection—which persisted for the rest of the monkey's life (2½ years).

Some further information is given about the distribution, cultivation, emetine-resistance, and other peculiarities of the forms of Endolimax found in Macaques and Man; and the findings of other workers with similar organisms are briefly considered. From all the evidence available it is concluded that E. nana naturally lives not only in Man but also in at least 3 different species of Macacus, and probably in monkeys of this genus generally. As yet there is no evidence that more than one species of Endolimax inhabits any of these hosts.

Infections with E. nana, whether in Man or Macaques, have usually been found to endure indefinitely; but a case (M. rhesus) has been observed in which the infection appears to have died out spontaneously after persisting for a known period of over 6 years.

Attempts to infect kittens with E. nana have all been unsuccessful hitherto, but there is evidence (from the work of others) that this amoeba may inhabit some other domestic mammals. Its host-distribution in nature needs further investigation.

Cytoadherence of Plasmodium falciparum-infected erythrocytes to melanoma cells was analysed using strains or isolates of parasites expressing or not expressing knobs (K+ or K− phenotype) on the erythrocyte surface. Both K+ and K− parasites had the capacity to cytoadhere to melanoma cells. Using a panel of melanoma cell lines with different surface expression of the cytoadherence receptors CD36, thrombospondin and ICAM-1 indicated that CD36 was the major receptor for parasites of both K+ and K− phenotypes. Binding competition experiments between K+ and K−-infected erythrocytes suggested that K+ cytoadherence is of higher affinity than that of K− parasites. However, some K− cytoadherence was also found in isolates containing mixed populations of K+ and K− parasites. The interaction of the two types of infected erythrocytes with melanoma cells also differed ultrastructurally, erythrocytes of K+ phenotype showing intimate interdigitations with microvilli on the melanoma cells, while erythrocytes of K− phenotype displayed more separated interactions with fewer sites of contact and involving only a few melanoma cell microvilli. One and the same infected erythrocyte may co-express the ligand for CD36-mediated cytoadherence and the structures mediating binding of uninfected erythrocytes to form rosettes.

In an attempt to find possible targets for benzimidazole action in muscle-stage larvae of Trichinella spiralis, the effects of mebendazole and thiabendazole were tested in vivo by oral treatment of infested mice and in vitro by including these anthelmintics in an adequate maintenance medium containing decapsulated larvae. The effects of the anthelmintics on succinate dehydrogenase and fumarate reductase activities, measured in the mitochondrial fraction obtained from the in vivo- or in vitro-treated larvae showed that only thiabendazole causes significant inhibition of fumarate reductase activity. On the other hand, measurements of free glucose, glycogen reserves and soluble protein in the treated larvae indicate that in vivo, mebendazole and thiabendazole clearly diminish free glucose levels, although in vitro only mebendazole produces the same diminution. Both the glycogen and protein contents of the larvae remained unchanged after treatment in vivo or in vitro. The importance of these findings with regard to a possible site of action for mebendazole and thiabendazole is discussed.

Severe adverse events (SAEs) following ivermectin treatment may occur in people harbouring high Loa loa microfilarial (mf) densities. In the context of mass ivermectin distribution for onchocerciasis control in Africa, it is crucial to define precisely the geographical distribution of L. loa in relation to that of Onchocerca volvulus and predict the prevalence of heavy infections. To this end, we analysed the distribution of mf loads in 4183 individuals living in 36 villages of central Cameroon. Mf loads were assessed quantitatively by calibrated blood smears, collected prior to ivermectin distribution. We explored the pattern of L. loa mf aggregation by fitting the (zero-truncated) negative binomial distribution and estimating its overdispersion parameter k by maximum likelihood. The value of k varied around 0·3 independently of mf intensity, host age, village and endemicity level. Based on these results, we developed a semi-empirical model to predict the prevalence of heavy L. loa mf loads in a community given its overall mf prevalence. If validated at the continental scale and linked to predictive spatial models of loiasis distribution, this approach would be particularly useful for optimizing the identification of areas at risk of SAEs and providing estimates of populations at risk in localities where L. loa and O. volvulus are co-endemic.

Adult specimens of Trochopus pini attach themselves by folding the disk-shaped haptor around the edge of the host's primary gill lamella. They are able to move from one place to another with the aid of disk-shaped pads on the head region.Ducts from the posterior median gland cells, which are extensive in the head region of the adult, open on the periphery of each disk and exude an adhesive secretion.

The oncomiracidium of T. pini is described and evidence is offered in support of direct invasion of the gills by these free-swimming larvae. Special attention is paid to the development of the head region of the parasite. Newly attached larvae lose their eye lenses and possibly their anterior median gland cells; their lateral head glands are well developed and most probably serve for attachment of the head region, since the posterior median gland cells are poorly developed and appear to have no ducts. Later the posterior median gland cells and their ducts increase in numbers and, in association with the pads on the head region, take over from the lateral glands the job of attaching the head region.

The paths of the tendons associated with the accessory sclerites on the haptor of T. pini indicate that this gill parasite has evolved relatively recently from the ancestors of benedeniid skin parasites.

I am most grateful to Commander J. Pareira, the Director of the Aquario Vasco da Gama in Lisbon where most of this work was done, and to the Staff of the Aquario and especially to Mr A. Ferreira. I am indebted to the Calouste Gulbenkian Foundation for a generous grant which made this visit possible. I am grateful also to the Directors and Staff of the Laboratoire Arago, Banyuls, France, the Station Biologique of the University of Bordeaux at Arcachon in France, and the Laboratory of the Marine Biological Association of the United Kingdom at Plymouth, England. I would like to thank the Royal Society for a grant from the Browne Fund which enabled me to work at Banyuls in 1962 and for a Pergamon Travel Award which made it possible to visit Arcachon in 1967.

The phylogenetic relationship of 5 genera, i.e. Diplozoon Nordmann, 1832, ParadiplozoonAchmerov, 1974, InustiatusKhotenovsky, 1978, SindiplozoonKhotenovsky, 1981, and EudiplozoonKhotenovsky, 1985 in the subfamily Diplozoinae Palombi, 1949 (Monogenea, Polyopisthocotylea) was inferred from rDNA ITS-2 region using neighbour-joining (NJ), maximum likelihood (ML) and Bayesian methods. The phylogenetic trees produced by using NJ, ML and Bayesian methods exhibit essentially the same topology. Surprisingly, freshwater species of Paradiplozoon from Europe clustered together with species of Diplozoon, but separated from Chinese Paradiplozoon species. The results of molecular phylogeny and lower level of divergence (4·1–15·7%) in ITS-2 rDNA among Paradiplozoon from Europe and Diplozoon and, on the other hand, high level of divergence (45·3–53·7%) among Paradiplozoon species from Europe and China might indicate the non-monophyletic origin of the genus Paradiplozoon. Also, the generic status of European Paradiplozoon needs to be revised. The species of Paradiplozoon in China is a basal group in Diplozoinae as revealed by NJ and Bayesian methods, and Sindiplozoon appears to be closely related to European Paradiplozoon and Diplozoon with their relationship to Eudiplozoon and Inustiatus being unresolved.

Chagas heart disease is a major public concern since 30% of infected patients develop cardiac alterations. The relationship between Trypanosoma cruzi discrete typing units (DTUs) and the biological properties exhibited by the parasite population has yet to be elucidated. In this study, we analysed the expression of α-smooth muscle actin (α-SMA) and galectin-3 (Gal-3) associated with cardiac extracellular matrix (ECM) remodelling a murine chronic cardiomyopathy induced by Tc I genotypes. We found the induction of myocarditis was associated with the upregulation of Col I, α-SMA, Gal-3, IFN-γ and IL-13, as analysed by q-PCR. In myocardial areas of fibrosis, the intensity of myocarditis and significant ECM remodelling correlated with the presence of Col I-, Gal-3- and α-SMA-positive cells. These results are promising for the further efforts to evaluate the relevance of Gal-3 in Chagas heart disease, since this galectin was proposed as a prognosis marker in heart failure patients.

The respiratory chain of the procyclic stage of Trypanosoma brucei contains the standard complexes I through IV, as well as several alternative enzymes contributing to electron flow. In this work, we studied the function of an alternative NADH : ubiquinone oxidoreductase (NDH2). Depletion of target mRNA was achieved using RNA interference (RNAi). In the non-induced and RNAi-induced cell growth, membrane potential change, alteration in production of reactive oxygen species, overall respiration, enzymatic activities of complexes I, III and/or IV and distribution of NADH : ubiquinone oxidoreductase activities in glycerol gradient fractions were measured. Finally, respiration using different substrates was tested on digitonin-permeabilized cells. The induced RNAi cell line exhibited slower growth, decreased mitochondrial membrane potential and lower sensitivity of respiration to inhibitors. Mitochondrial glycerol-3-phosphate dehydrogenase was the only enzymatic activity that has significantly changed in the interfered cells. This elevation as well as a decrease of respiration using NADH was confirmed on digitonin-permeabilized cells. The data presented here together with previously published findings on complex I led us to propose that NDH2 is the major NADH : ubiquinone oxidoreductase responsible for cytosolic and not for mitochondrial NAD+ regeneration in the mitochondrion of procyclic T. brucei.

Entamoeba histolytica is an enteric parasite that infects approximately 50 million people worldwide. Although E. histolytica is a zoonotic parasite that has the potential to infect nonhuman primates, such transmission is poorly understood. Consequently, this study examined whether E. histolytica is present among humans, chimpanzees and baboons living in the Greater Gombe Ecosystem (GGE), Tanzania. The primary aims were to determine patterns of E. histolytica infection in a system with human-nonhuman primate overlap and to test associations between infection status and potential risk factors of disease. Entamoeba spp. occurred in 60.3% of human, 65.6% of chimpanzee and 88.6% of baboon samples. Entamoeba histolytica occurred in 12.1% of human, 34.1% of chimpanzee and 10.9% of baboon samples. Human E. histolytica infection was associated with gastrointestinal symptoms. This was the first study to confirm the presence of E. histolytica in the GGE. The high sample prevalence of E. histolytica in three sympatric primates suggests that zoonotic transmission is possible and stresses the need for further phylogenetic studies. Interventions targeting better sanitation and hygiene practices for humans living in the GGE can help prevent E. histolytica infection in humans, while also protecting the endangered chimpanzees and other primates in this region.

Carbon dioxide (CO2) is an important sensory cue for many animals, including both parasitic and free-living nematodes. Many nematodes show context-dependent, experience-dependent and/or life-stage-dependent behavioural responses to CO2, suggesting that CO2 plays crucial roles throughout the nematode life cycle in multiple ethological contexts. Nematodes also show a wide range of physiological responses to CO2. Here, we review the diverse responses of parasitic and free-living nematodes to CO2. We also discuss the molecular, cellular and neural circuit mechanisms that mediate CO2 detection in nematodes, and that drive context-dependent and experience-dependent responses of nematodes to CO2.

Chagas disease is one of the most important vector-borne zoonotic diseases in Latin America. Control strategies could be improved if transmissibility patterns of its aetiologic agent, Trypanosoma cruzi, were better understood. To understand transmissibility patterns of Chagas disease in Mexico, we inferred potential vectors and hosts of T. cruzi from geographic distributions of nine species of Triatominae and 396 wild mammal species, respectively. The most probable vectors and hosts of T. cruzi were represented in a Complex Inference Network, from which we formulated a predictive model and several associated hypotheses about the ecological epidemiology of Chagas disease. We compiled a list of confirmed mammal hosts to test our hypotheses. Our tests allowed us to predict the most important potential hosts of T. cruzi and to validate the model showing that the confirmed hosts were those predicted to be the most important hosts. We were also able to predict differences in the transmissibility of T. cruzi among triatomine species from spatial data. We hope our findings help drive efforts for future experimental studies.

This study examines polymorphisms in 2 genes (pfmdr1 and cg2), which have been associated with resistance to chloroquine in Plasmodium falciparum, to determine their value as predictors of resistance status. Among field samples from children in Zaria, northern Nigeria, the Tyr-86 polymorphism in pfmdr1 and Ala-281 and the Dd2κ repeat of cg2, were significantly associated. In 8 samples classified resistant by the micro-in vitro test, or, where this failed, by in vivo trial, 7 showed the cg2 Dd2 type κ repeat, and 6 of these had both the Ala-281 allele and the pfmdr1 Tyr-86 allele. In 26 chloroquine-sensitive samples, none had this combination of 3 polymorphisms (P = 0·00002). This indicates 75% sensitivity and 100% specificity in detection of resistance and shows a positive predictive value for resistant infections of 100%. The negative predictive value, because of sensitivity less than 100%, would depend on the prevalence of resistance. Where prevalence of resistance is approx. 21% as in Zaria, the negative predictive value would be 94%, while in Gabon, with a prevalence of ca 73% it would be 60%. The use of (cg2: Ala-281, Dd2κ. pfmdr1: Tyr-86) genotype detection as a predictive epidemiological tool to examine the distribution of chloroquine-resistance in parts of Africa is therefore possible. The sensitivity of detection of resistant strains still requires improvement.