This paper generalizes the two-dimensional approximation of models of macroparasites on homogeneous populations developed by Anderson & May (1978), focusing on how the dispersion (the variance to mean ratio) of the equilibrium distribution of parasites on hosts is related to the stability of the equilibrium. We show in the approximate system that the equilibrium is stabilized not by aggregation, but by dispersion which increases as a function of the mean. Computer simulations indicate, however, that this analysis fails to capture properly the dynamics of the full system, raising the question of whether any two-dimensional system could produce an adequate approximation. We discuss the relevance of our results to several empirical studies which have examined the relation of dispersion to the mean.

The Paramyxida, closely related to haplosporidians, paradinids, and mikrocytids, is an obscure order of parasitic protists within the class Ascetosporea. All characterized ascetosporeans are parasites of invertebrate hosts, including molluscs, crustaceans and polychaetes. Representatives of the genus Marteilia are the best studied paramyxids, largely due to their impact on cultured oyster stocks, and their listing in international legislative frameworks. Although several examples of microsporidian hyperparasitism of paramyxids have been reported, phylogenetic data for these taxa are lacking. Recently, a microsporidian parasite was described infecting the paramyxid Marteilia cochillia, a serious pathogen of European cockles. In the current study, we investigated the phylogeny of the microsporidian hyperparasite infecting M. cochillia in cockles and, a further hyperparasite, Unikaryon legeri infecting the digenean Meiogymnophallus minutus, also in cockles. We show that rather than representing basally branching taxa in the increasingly replete Cryptomycota/Rozellomycota outgroup (containing taxa such as Mitosporidium and Paramicrosoridium), these hyperparasites instead group with other known microsporidian parasites infecting aquatic crustaceans. In doing so, we erect a new genus and species (Hyperspora aquatica n. gn., n.sp.) to contain the hyperparasite of M. cochillia and clarify the phylogenetic position of U. legeri. We propose that in both cases, hyperparasitism may provide a strategy for the vectoring of microsporidians between hosts of different trophic status (e.g. molluscs to crustaceans) within aquatic systems. In particular, we propose that the paramyxid hyperparasite H. aquatica may eventually be detected as a parasite of marine crustaceans. The potential route of transmission of the microsporidian between the paramyxid (in its host cockle) to crustaceans, and, the ‘hitch-hiking’ strategy employed by H. aquatica is discussed.

We studied variation in the structure of component communities of helminths in sand lizards, Lacerta agilis, from 30 localities in the Ukraine and Bulgaria. Thirty-five separate samples of lizards, with a total of 661 completely censused infracommunities, yielded 30 helminth species (4 cestodes, 10 trematodes, 3 acanthocephalans and 13 nematodes). In its range within the Ukraine, L. agilis serves as the final host for 13 species of which only 3 (S. lacertae, S. hoffmanni and P. molini) can be considered as lizard specialists. A characteristic feature of these helminth component communities was the large proportion of heteroxeneous helminth species for which L. agilis serves as paratenic host. Sand lizards in the meadow steppeland zone were primarily parasitized by larval helminths that represented a major proportion of the total number of all worms recovered while those sampled in the grassland/forest transition zone were characterized by substantially higher proportions of adult helminths using lizards as final hosts. However, L. agilis was parasitized by a much higher proportion of lizard specialists in the ‘typical' habitats of the meadow steppeland zone as opposed to those located in ‘marginal' habitats in the grassland/forest transition zone, where helminths were shared to a greater extent with amphibian hosts.

Groupers (Epinephelinae) are prominent marine fishes distributed in the warmer waters of the world. Review of the literature suggests that trematodes are known from only 62 of the 159 species and only 9 of 15 genera; nearly 90% of host–parasite combinations have been reported only once or twice. All 20 families and all but 7 of 76 genera of trematodes found in epinephelines also occur in non-epinephelines. Only 12 genera of trematodes are reported from both the Atlantic–Eastern Pacific and the Indo–West Pacific. Few (perhaps no) species are credibly cosmopolitan but some have wide distributions across the Indo–West Pacific. The hierarchical ‘relatedness’ of epinephelines as suggested by how they share trematode taxa (families, genera, species) shows little congruence with what is known of their phylogeny. The major determinant of relatedness appears to be geographical proximity. Together these attributes suggest that host-parasite co-evolution has contributed little to the evolution of trematode communities of epinephelines. Instead, they appear to have arisen through localized episodes of host-switching, presumably both into and out of the epinephelines. The Epinephelinae may well be typical of most groups of marine fishes both in the extent to which their trematode parasites are known and in that, apparently, co-evolution has contributed little to the evolution of their communities of trematodes.

Several monoclonal antibodies (Mabs) were raised against the L1 muscle stage (L1M) of Trichinella spiralis (Ts) and Trichinella pseudospiralis (Tp). Western blot analysis of various antigenic preparations established that Mabs described by different authors recognized 8 antigenic fractions (TSL1–TSL8) in crude extracts of infective larvae. The TSL1 fraction was immunodominant and present on the cuticle of different parasite stages. Mabs against Trichinella T5 (T5) and Ts were selected in order to extend the previous studies to another Trichinella phenotype. Only 35% of the selected Mabs recognized linear epitopes and 71% reacted with soluble or excretory–secretory antigens in a dot blot procedure and ELISA test. The targets of the Mabs were identified by immunoprecipitation with [35S]methionine-labelled L1M worm lysate. Mabs prepared from mice immunized with the whole parasite (T5) recognized a wider panel of antigens in different parasitic organs. Seven antigenic structures were distinguished on the cuticle and several epitopes were identified in the gut, haemolymph and stichocytes. Eleven antigenic groups were established according to their indirect immunofluorescence pattern on cross-sections of the worm. Monoclonal antibodies raised against Ts soluble antigen mainly recognized epitopes in stichocytes and on the cuticle surface. All the selected Mabs recognized T5 and Trichinella britovi (Tb) strengthening the link between these 2 species. Four Mabs were used to differentiate antigenic structures among 6 Trichinella phenotypes and to develop a new tool to follow gene flow within the Trichinella genus.

Light microscopy of stained blood films is still the mainstay of malaria diagnosis in many regions, but its pre-eminence is threatened by accurate and sensitive rapid diagnostic tests (RDTs) based on immunochromatography which are now widely used in the field. In well-resourced regions, nucleic acid detection easily out performs microscopy and RDTs. This paper reviews the main in vitro methods for parasite detection and considers future trends in diagnostics, both for sophisticated laboratory settings and for field use.

Free-living animals are usually inhabited by a community of parasitic species that can interact with each other and alter both host susceptibility and parasite transmission. In this study we tested the prediction that an increase in the gastrointestinal nematode Heligmosomoides polygyrus would increase the infestation of the tick Ixodes ricinus, in free-living yellow-necked mice, Apodemus flavicollis. An extensive cross-sectional trapping survey identified a negative relationship between H. polygyrus and I. ricinus counter to the prediction. An experimental reduction of the nematode infection through anthelmintic treatment resulted in an increase in tick infestation, suggesting that this negative association was one of cause and effect. Host characteristics (breeding condition and age) and habitat variables also contributed to affect tick infestation. While these results were counter to the prediction, they still support the hypothesis that interactions between parasite species can shape parasite community dynamics in natural systems. Laboratory models may act differently from natural populations and the mechanism generating the negative association is discussed.

Nippostrongylus brasiliensis induces a biphasic anorexia in laboratory rats, the first phase coincident with lung invasion (ca day 2) and the second when the worms mature in the intestine (ca day 8). Using the anthelminthic, mebendazole (MBZ), N. brasiliensis infections of the rat were eliminated between the first and second anorexic episodes. This intervention prevented the expression of the second phase of anorexia. Rats exposed to a second infection with N. brasiliensis, 3 weeks after the primary infection, exhibited only a first phase anorexic response which was not influenced by MBZ termination of the primary infection. The lower cumulative food intake and weight gain of all infected rats after 8 days of infection were accompanied by elevated plasma insulin and, in some individuals, by elevated plasma leptin, compared with uninfected controls and previously-infected MBZ-treated rats. Messenger RNA levels for neuropeptide Y were higher in the hypothalamic arcuate nucleus of 8-day infected rats than in recovering MBZ-treated animals. Inoculation of rats with heat-killed N. brasiliensis larvae failed to induce anorexia and did not alter the severity of biphasic anorexia on subsequent injection of viable larvae. The first anorexic episode is therefore dependent upon viable migrating larvae. The second phase of anorexia clearly requires the continuing presence of the parasite beyond the lung phase. Viable migrating larvae are also required to confer ‘resistance’ to reinfection.

The attachment of Schistosoma mansoni cercariae to mammalian skin is specifically stimulated by L-arginine. As L-arginine is an unsuitable signal for a specific identification of mammalian skin we examined the following 5 hypotheses to explain the advantage of the cercarial sensitivity to L-arginine. (1) A Schistosoma infection lowered the arginine concentration in the serum of mice, and this could enable the cercariae to avoid attachments to already infected mice. However, the infection did not reduce the arginine concentration in the skin and the cercarial attachment responses to it. (2) Creeping cercariae showed chemotactic orientation specifically along increasing L-arginine gradients. L-arginine could act as a pheromone which could guide cercariae towards common penetration sites. However, the cercarial acetabular gland contents were not attractive and they did not (in contrast to previous reports) contain much arginine. (3) Schistosomula (transformed cercariae) could use L-arginine to produce nitric oxide (NO) for blood vessel dilation during their migration in the host. However, in vitro the transformed cercariae did not convert L-arginine into citrulline and NO. (4) Schistosomula could bind L-arginine from the surrounding tissues and so escape the cellular immune attack (which needs L-arginine as the precursor of NO). However, transformed cercariae bound no more L-arginine than L-serine and L-lysine. (5) Schistosomula, migrating parallel to the surface in the mammalian epidermis, are dependent on information on their position between the inner and the surface layers of the skin. In the mouse skin, they adjusted their body axis with the ventral side toward the deeper (arginine-residue rich) epidermis layers. When migrating in agar, they showed chemo-orientation toward serum, and D-glucose and L-arginine were the stimulating compounds therein. The burrowing schistosomula adjusted their body axis (as in the epidermis) with the ventral side toward the higher concentration of L-arginine and not of glucose. We argue that the sensitivity for L-arginine has its primary function in orientation within mammalian skin and in location of blood vessels.

We applied a combined molecular and morphological approach to resolve the taxonomic status of Saccocoelium spp. parasitizing sympatric mullets (Mugilidae) in the Mediterranean. Eight morphotypes of Saccocoelium were distinguished by means of multivariate statistical analyses: 2 of Saccocoelium obesum ex Liza spp.; 4 of S. tensum ex Liza spp.; and 2 (S. cephali and Saccocoelium sp.) ex Mugil cephalus. Sequences of the 28S and ITS2 rRNA gene regions were obtained for a total of 21 isolates of these morphotypes. Combining sequence data analysis with a detailed morphological and multivariate morphometric study of the specimens allowed the demonstration of cryptic diversity thus rejecting the hypothesis of a single species of Saccocoelium infecting sympatric mullets in the Mediterranean. Comparative sequence analysis revealed 4 unique genotypes, thus corroborating the distinct species status of Saccocoelium obesum, S. tensum and S. cephali and a new cryptic species ex Liza aurata and L. saliens recognized by its consistent morphological differentiation and genetic divergence. However, in spite of their sharp morphological difference the 2 morphotypes from M. cephalus showed no molecular differentiation and 4 morphotypes of S. tensum were genetically identical. This wide intraspecific morphological variation within S. tensum and S. cephali suggests that delimiting species of Saccocoelium using solely morphological criteria will be misleading.

1. Crepidostomum farionis (Müller, 1784) and C. metoecus (Braun, 1900) were found to occur together in the same host fish, Salmo trutta L., S. salar L. and Anguilla anguilla (L.) in mid-Wales. It is believed that the latter species constituted an accidental host.

2. C. metoecus is recorded in Britain for the first time.

3. There was some evidence of a habitat isolation in individual fish as C. metoecus occurred predominantly within the pyloric caeca, while C. farionis usually occupied a more posterior station in the intestine.

4. A detailed description is given of G. metoecus and the salient features of this species are compared with those of C. farionis.

5. The two species of Crepidostomum commonly occurred in large numbers in their fish hosts, individual fish harbouring up to 157 worms. No lesions attributable to Crepidostomum were, however, detected and there was no evidence of pathogenicity.

6. It would appear that the eel is physiologically immune to both species of Crepidostomum and that the salmon parr is partially resistant.

7. The degree of infestation of trout and parr with both the Crepidostomum species is at a maximum during the winter months and at a minimum in the summer months. This seasonal variation can be attributed to a periodicity in the swarming of the cercariae.

8. There is no evidence of age resistance or acquired immunity of the trout to infestation by Crepidostomum species.

Numerical analyses of Trypanozoon taxonomy are presented, based on the isoenzyme data of Stevens et al. (1992). The previous study used a reduced range of enzymes compared with earlier work; the analyses indicate the value of this rationalized system. Both recently isolated trypanosome stocks and previously studied populations were included, allowing detailed comparison with earlier studies. Relationships between zymodemes were calculated with an improved similarity coefficient program, using Jaccard's coefficient (1908), and by Nei's method (1972). Dendrograms were constructed from the matrices produced with the group-average method. The groupings produced by both numerical methods were in close agreement, and the clusters of related principal zymodemes largely matched the species, subspecies and strain groups proposed by previous workers. Trypanozoon biochemical taxonomy is reviewed and the groupings reinforced by this study are: the mainly East African strain groups, busoga, zambezi, kakumbi, kiboko and sindo; T.b. gambiense and the bouaflé strain group from West Africa, and T. evansi; an intermediate bouaflé/busoga group was also recognized.

A monoclonal antibody, 1C4, was produced which recognizes a 65 kDa protein that is localized to the plasma membrane of human erythrocytes infected with Plasmodium falciparum. By immunofluorescence the antigen was visualized as dots on the surface of the infected cell. The 65 kDa protein was present in 4 strains of diverse geographical origin, and in erythrocytes infected with a knobless strain. The 65 kDa protein was insoluble in non-ionic detergents, but was partly soluble in SDS and some high (1 M) śalt solutions. The 65 kDa protein is recognized by antibodies specific for the cytoplasmic domain and the N-terminal side of the membrane-spanning region of human band 3, but was not recognized by an antibody specific to the C-terminal side of the membrane-spanning region. The results of treatment of the 65 kDa protein with trypsin and chymotrypsin are consistent with the 65 kDa protein being a truncated and covalently modified band 3 molecule which consists of the first 540 amino acids of human band 3.

The sexual cycle of Theileria taurotragi was identified in the gut lumen of replete Rhipicephalus appendiculatus nymphs maintained at 28°C which had fed on eland with rising parasitaemias. Macro- and microgametes developed from ring-form piroplasms within 24 h after repletion. The microgamonts were elongate with 2 or more lateral projections. The nuclei of the microgamonts divided into 4 and the microgamonts differentiated into 4 thread-like microgametes each with a central nucleus. Round macrogametes developed at the same time and stages indicative of the fusion of the macro- and microgametes were observed after 48 h. The resultant zygotes were detected in the gut wall cells by day 4, many being rounded and vacuolated with a peripheral nucleus. By day 7 the zygote cytoplasm became dense and they lay in clusters still within the cells. Binucleate zygotes were observed at this stage. The zygotes increased in size and by day 12 began to transform into kinetes by invagination. By the time the nymphs moulted into adults (about day 14), the kinete straightened to a broad anterior end and a tapering posterior with a mean length of 22·1 μm. Kinetes were detected in the haemolymph by day 16. By day 20 the kinetes had penetrated the salivary gland acinar cells where they underwent schizogony until the infected acinar cells were filled with multinucleated sporoblasts. The nuclei of the infected acinar cells became greatly enlarged soon after penetration of the kinete. Division of the sporoblast nuclei was stimulated by feeding of the adult ticks. From the 2nd day of attachment of the ticks to rabbits the sporoblasts underwent a process of schizogony to produce cytomeres. Each nucleus of the cytomere divided to produce several small nuclei which differentiated into uninucleated sporozoites.

When incubated at 13°C in alternating 12 h periods of light and darkness eggs of Acanthocotyle lobianchi rarely hatch. Hatching was not induced by shadowing or by mechanical disturbance. When skin mucus from the host ray was added to eggs which were more than 15 days old, hatching occurred almost immediately in about 60% of all eggs tested. Susceptibility to host mucus increased with the age of the eggs so that after 26–30 days of incubation over 80% of the eggs tested hatched. Some oncomiracidia were found to remain alive for up to 83 days in the egg, although they appeared to be fully developed at 15 days. Lipid droplets in the body of the 15-day-old oncomiracidia gradually disappeared as the eggs aged and it is suggested that these droplets serve as a food store for resting larvae.

Random amplified polymorphic DNA markers (RAPD) were used to visualize the genetic diversity within and between infrapopulations of Schistosoma mansoni recovered from the natural vertebrate host, Rattus rattus, trapped at an insular Guadeloupean focus. Phenotypes were characterized by the sex of the parasites and by 8 polymorphic markers generated by 3 primers. Among the 212 parasite individuals recovered from 10 infected rats, 78 genotypes were characterized. All the hosts naturally infected harboured multiple parasite genotypes with a maximum diversity of 28 genotypes/host. Phenotypic and genotypic diversity calculated by Shannon-Wiener's indices and Lynch and Milligan's estimators respectively is, on average, greater within than between hosts. Considering the very low snail infection rates observed in this focus and the rapid turnover of the vertebrate hosts, our results suggest that the high mobility of the vertebrate host and/or plurimiracidial snail infections could be factors responsible for parasite genetic diversity within hosts.

The post-genomics era has provided researchers with access to a new generation of tools for the global characterization and understanding of pathogen diversity. This review provides a critical summary of published Leishmania post-genomic research efforts to date, and discusses the potential impact of the addition of metabolomics to the post-genomic toolbox. Metabolomics aims at understanding biology by comprehensive metabolite profiling. We present an overview of the design and interpretation of metabolomics experiments in the context of Leishmania research. Sample preparation, measurement techniques, and bioinformatics analysis of the generated complex datasets are discussed in detail. To illustrate the concepts and the expected results of metabolomics analyses, we also present an overview of comparative metabolic profiles of drug-sensitive and drug-resistant Leishmania donovani clinical isolates.

X-ray micro-computed tomography (μCT) is a technique which can obtain three-dimensional images of a sample, including its internal structure, without the need for destructive sectioning. Here, we review the capability of the technique and examine its potential to provide novel insights into the lifestyles of parasites embedded within host tissue. The current capabilities and limitations of the technology in producing contrast in soft tissues are discussed, as well as the potential solutions for parasitologists looking to apply this technique. We present example images of the mouse whipworm Trichuris muris and discuss the application of μCT to provide unique insights into parasite behaviour and pathology, which are inaccessible to other imaging modalities.

The objective of this experiment was to investigate a possible interaction between protein and energy malnutrition (PEM) and intestinal nematode infections. We report on a 3×2 factorial study in which pigs were fed either a low protein energy (LPE), low protein (LP) or a normal protein energy (NPE) diet, and 6 weeks later inoculated with Trichuris suis (4000 infective eggs). Secondarily, in order to obtain a polyparasitic status, pigs were concomitantly inoculated with Ascaris suum (600 infective eggs). The number of T. suis-infected pigs was higher in LP pigs compared with NPE pigs (100 versus 58%; P = 0·037), although the differences in median T. suis worm burdens between groups at necropsy 10 weeks post-infection (p.i.) (LPE: 795; LP: 835; NPE: 48 worms; P = 0·33) were not significant. Interestingly, only T. suis in NPE were highly aggregated (k = 0·44), in contrast to a more uniform distribution among pigs in LPE (k = 1·43) and LP (k = 1·55) i.e. the majority of pigs harboured moderate worm burdens in LPE and LP, while most pigs had few or no worms in NPE. Further, T. suis worms in the LPE and LP groups were decreased in length (mean: LPE: 23·5 mm; LP: 24·3 mm; NPE: 29·4 mm; P = 0·004). The pre-patency period of T. suis was also extended in the LPE and LP groups, as reflected by lower faecal egg output at week 6 (P = 0·048) and/or 7 p.i. (P = 0·007). More A. suum worms were recovered from LP compared with the NPE group (mean: 5·4 versus 0·6; P = 0·040); this was accompanied by a higher faecal egg output in the former (P = 0·004). The low protein diets resulted in lower pig body weight gains, serum albumin, haemoglobin and packed cell volume (PCV) levels as well as diminished peripheral eosinophil counts. Infection significantly altered these parameters in the low protein groups, i.e. the pathophysiological consequences of infection were more severe in the PEM pigs. These results demonstrate that reduced protein in the diet leads to malnourishment of both the host pigs and T. suis, and compromises the pig's ability to resist infection by T. suis and A. suum.

Although bats are one of the most successful and diverse of mammalian orders, studies that focus upon bat endoparasites are limited. To further knowledge of bat parasitology, pipistrelle bats (Pipistrellus pipistrellus and P. pygmaeus) were acquired from across the Greater Manchester and Lancashire region of England and examined for gastrointestinal helminths using morphological and molecular analyses†. Sixty-eight of 90 adult/juvenile bats (76% prevalence) were infected with at least 1 species of helminth and mean helminth abundance was 48·2 (+/−7·0). All helminths were digenean trematodes and the following species were identified in 51 P. pipistrellus specimens (prevalence in parentheses): Lecithodendrium linstowi (80·4%), L. spathulatum (19·6%), Prosthodendrium sp. (35·3%), Plagiorchis koreanus (29·4%) and Pycnoporus heteroporus (9·8%). Statistical analyses, incorporating multifactorial models, showed that male bats exhibited a significantly more aggregated helminth distribution and lower abundance than females. Positive associations were observed between L. linstowi and L. spathulatum, Prosthodendrium sp. and P. heteroporus and between L. spathulatum and P. koreanus. A revised phylogeny of bat-associated Lecithodendriidae, incorporating novel L. spathulatum and Prosthodendrium sp. 28S rRNA sequences, separated the controversial clade formed by L. linstowi and P. hurkovaae. Further studies are likely to assist the understanding of bat-parasite/pathogen relationships, helminth infracommunity structures and phylogenetics.