The multifaceted interactions occurring between gastrointestinal (GI) parasitic helminths and the host gut microbiota are emerging as a key area of study within the broader research domain of host-pathogen relationships. Over the past few years, a wealth of investigations has demonstrated that GI helminths interact with the host gut flora, and that such interactions result in modifications of the host immune and metabolic statuses. Nevertheless, whilst selected changes in gut microbial composition are consistently observed in response to GI helminth infections across several host-parasite systems, research in this area to date is largely characterised by inconsistent findings. These discrepancies are particularly evident when data from studies of GI helminth-microbiota interactions conducted in humans from parasite-endemic regions are compared. In this review, we provide an overview of the main sources of variance that affect investigations on helminth-gut microbiota interactions in humans, and propose a series of methodological approaches that, whilst accounting for the inevitable constraints of fieldwork, are aimed at minimising confounding factors and draw biologically meaningful interpretations from highly variable datasets.

Target-based approaches for human African trypanosomiasis (HAT) and related parasites can be a valuable route for drug discovery for these diseases. However, care needs to be taken in selection of both the actual drug target and the chemical matter that is developed. In this article, potential criteria to aid target selection are described. Then the physiochemical properties of typical oral drugs are discussed and compared to those of known anti-parasitics.

It recently became clear that adult eo- and palaeacanthocephalans parasitizing fish can bioconcentrate several heavy metals to significantly higher concentrations than the tissues of their definitive hosts. Following this discovery the lead accumulation of the archiacanthocephalan Moniliformis moniliformis was investigated using experimentally infected male Wistar rats of the CD-M-strain. The worms were allowed to grow up for 4 weeks post-infection followed by a 3 weeks oral lead exposure of the rats. After the exposure period the rats were killed and the metal levels were determined in muscle, liver, intestine and kidney of the rats as well as in different organs of female and male acanthocephalans. Lead concentrations were found to be highest in female M. moniliformis followed by the kidneys of the rats. Male worms contained approximately the same lead concentration as the hosts' kidneys. Lead analysis of the worms' organs revealed the highest lead concentration in the eggs of female acanthocephalans, followed by the cement gland of male Worms. Whilst the lead burden of the presoma was higher than that detected in the kidneys of the rats, the lead content of the metasoma was even lower than in the kidneys. A lead uptake of M. moniliformis from the intestinal lumen of the host became apparent as the faeces of infected rats contained significantly less lead compared to the uninfected conspecifics. Thus, this study reveals that lead accumulation also occurs in archiacanthocephalans parasitizing mammals. But the degree of metal bioconcentration is considerably lower compared to eo- and palaeacanthocephalans in fish. Anyway, due to a lack of adequate sentinel species in terrestrial biotopes the host–parasite system rat– M. moniliformis appears to be a useful and promising bioindication system especially in urban ecosystems in temperate regions.

Patterns of infection among hosts in a population are often driven by intrinsic host features such as age or sex, as well as by positive or negative interactions between parasite species. We investigated helminth parasitism in 2 South American rodent species, Ctenomys australis and C. talarum (Octodontidae), to determine whether the unusual solitary and subterranean nature of these hosts would impact their patterns of infection. We applied generalized linear models to infection data on a total of 7 helminth species (1 in C. australis and 6 in C. talarum). Host age and season of capture influenced infection levels in some of the helminth species, but none were influenced by host body condition. In C. talarum, 4 pairs of helminth species showed significant associations, either asymmetrical or symmetrical, and with 3 of the 4 being positive; strong inter-specific facilitation appears likely in 1 case. Also, we found that female hosts, especially non-pregnant ones, harboured heavier infections of 2 nematode species than male hosts. This is in sharp contrast to the general male-bias reported for most studies of nematodes in wild mammals, and we develop explanations for these results based on the unusual ecology of these subterranean rodents.

A cDNA-AFLP (cDNA-Amplified Fragment Length Polymorphism)-based strategy has been used to identify levamisole (LEV) resistance markers in the nematode Haemonchus contortus. Transcript profiles of adult nematodes from two LEV-resistant and two susceptible isolates were compared. Among the 17 280 transcript-derived fragments (TDFs) amplified, 26 presented a polymorphic pattern between resistant and susceptible nematodes: 11 TDFs were present in both resistant isolates and absent from both susceptible isolates whereas 15 TDFs were present in both susceptible isolates and absent from both resistant isolates. 8 TDFs specifically present in resistant isolates were cloned and sequenced. Some of these TDFs could represent novel genes, as their sequences presented no homologies in databases. Interestingly, specific expression of one candidate (HA17) in resistant nematodes from different isolates was confirmed by RT-PCR experiments. The finding that HA17 expression correlates with LEV resistance in three H. contortus isolates vs five susceptible isolates strongly suggest that we identified a new potential marker of LEV resistance. This differential approach at the transcriptome level could be of great interest for the identification of the molecular mechanism involved in this phenotype.

The levels of superoxide dismutase (SOD) were determined in detergent-soluble, somatic and excretion–secretion (E–S) preparations from adult Fasciola hepatica using the xanthine oxidase system and visualized in substrate gels. Compared to detergent-soluble and somatic extracts, E–S products showed the highest SOD activity (88 ·5 U/mg), indicating active release to the medium in which parasites were maintained. SOD specific activity was also detected at high levels in E–S products from 3-week-old and 5-week-old immature migrating flukes (25 and 143 U/mg, respectively). In all preparations except for the somatic extract, the activity was characterized as cyanide-sensitive CuZn SOD. Differences in SOD isoenzyme profiles between the extracts were observed in native polyacrylamide gel electrophoresis: the somatic and detergent-soluble extracts exhibited 1 band of activity while the E–S products from immature and adults flukes contained 2 and 3 migrating bands, respectively. SOD was purified from the detergent-soluble extract and E–S products of adult worms by a combination of ultrafiltration, gel filtration on Sephacryl S-200 HR and ion-exchange chromatography on QAE Sephadex A-50. The SOD from detergent-soluble extract showed, by SDS–PAGE analysis, 1 band of 16 kDa apparent molecular weight. The SOD from E–S products showed 2 bands of 16 and 60 kDa apparent molecular weight. N-terminal sequence analysis of the 16 kDa band from the detergent-soluble preparation showed some similarity with Schistosoma mansoni cytoplasmic SOD. These enzymes may have a potential role in the evasion of the oxidative burst killing mechanism by immune cells.

Bird-specific ticks do not infest humans and livestock, but these ticks often share their avian hosts with generalist ticks that do. Therefore, their feeding activity may have an impact on the transmission of pathogens outside bird–tick transmission cycles. Here we examined the seasonal feeding activity of the tree-hole tick (Ixodes arboricola) in relation to the activity of its hole-breeding hosts (Parus major and Cyanistes caeruleus). We analysed data on ticks derived from birds, on the abundance of engorged ticks inside nest boxes, and on bird nests that were experimentally exposed to ticks. We observed a non-random pattern of feeding associated with the tick instar and host age. The majority of adult ticks fed on nestlings, while nymphs and larvae fed on both free-flying birds and nestlings. Due to their fast development, some ticks were able to feed twice within the same breeding season. The highest infestation rates in free-flying birds were found during the pre-breeding period and during autumn and winter when birds roost inside cavities. Except during winter, feeding of I. arboricola overlapped in time with the generalist Ixodes ricinus, implying that tick-borne microorganisms that are maintained by I. arboricola and birds could be bridged by I. ricinus to other hosts.

The crayfish plague pathogen, Aphanomyces astaci, is one of the most serious threats to indigenous European crayfish species. The North American invasive spiny-cheek crayfish, Orconectes limosus, is an important source of this pathogen in central and western Europe. We evaluated potential changes in A. astaci spore release rate from infected individuals of this species by experiments investigating the pathogen transmission to susceptible noble crayfish, Astacus astacus. We filtered defined volumes of water regularly to quantify spore concentration, and sampled crayfish tissues at the end of the experiment. The filters and tissues were then tested for the presence of A. astaci DNA by species-specific quantitative PCR. Additionally, we tested the efficiency of horizontal transmission to apparently uninfected O. limosus. The experiments confirmed that A. astaci can be transmitted to susceptible crayfish during intermoult periods, and that the pathogen was more frequently detected in noble crayfish recipients than in American ones. The pathogen spore concentrations substantially varied in time, and significantly increased during moulting of infected hosts. Our study strengthens the evidence that although the likelihood of crayfish plague transmission by water transfer from localities with infected American crayfish might increase when these are moulting or dying, no time-periods can be proclaimed safe.

Many invertebrates are either parasites themselves or vectors involved in parasite transmission; thereby, the interactions of parasites with final or intermediate hosts are often mediated by glycans. Therefore, it is of interest to compare the glycan structures or motifs present across invertebrate species. While a typical vertebrate modification such as sialic acid is rare in lower animals, antennal and core modifications of N-glycans are highly varied and range from core fucose, galactosylated fucose, fucosylated galactose, methyl groups, glucuronic acid and sulphate through to addition of zwitterionic moieties (phosphorylcholine, phosphoethanolamine and aminoethylphosphonate). Only in some cases are the enzymatic bases and the biological function of these modifications known. We are indeed still in the phase of discovering invertebrate glycomes primarily using mass spectrometry, but molecular biology and microarraying techniques are complementary to the determination of novel glycan structures and their functions.

Species of Plasmodium (Plasmodiidae, Haemosporida) are widespread and cause malaria, which can be severe in avian hosts. Molecular markers are essential to detect and identify parasites, but still absent for many avian malaria and related haemosporidian species. Here, we provide first molecular characterization of Plasmodium matutinum, a common agent of avian malaria. This parasite was isolated from a naturally infected thrush nightingale Luscinia luscinia (Muscicapidae). Fragments of mitochondrial, apicoplast and nuclear genomes were obtained. Domestic canaries Serinus canaria were susceptible after inoculation of infected blood, and the long-lasting light parasitemia developed in two exposed birds. Clinical signs of illness were not reported. Illustrations of blood stages of P. matutinum (pLINN1) are given, and phylogenetic analysis identified the closely related avian Plasmodium species. The phylogeny based on partial cytochrome b (cyt b) sequences suggests that this parasite is most closely related to Plasmodium tejerai (cyt b lineage pSPMAG01), a common malaria parasite of American birds. Both these parasites belong to subgenus Haemamoeba, and their blood stages are similar morphologically, particularly due to marked vacuolization of the cytoplasm in growing erythrocytic meronts. Molecular data show that transmission of P. matutinum (pLINN1) occurs broadly in the Holarctic, and the parasite likely is of cosmopolitan distribution. Passeriform birds and Culex mosquitoes are common hosts. This study provides first molecular markers for detection of P. matutinum.

Factors constraining the evolution of host-specificity were investigated using a gastrointestinal parasitic nematode, Strongyloides ratti. S. ratti is a natural parasite of rats which can also reproduce, with decreased success, in laboratory mice. Observed host-specificity arose from lower establishment, reduced per capita fecundity and more rapid expulsion of parasites from mice relative to rats. Variation in the efficacy of thymus-dependent immunity between host species (rats and mice) was insufficient to explain the majority of the observed differences in parasite establishment and reproductive success. The role of natural selection in determining host-specificity was addressed using experimental selection followed by reciprocal fitness assays in both host species. Experimental selection failed to modify the host-specificity of S. ratti to any measurable degree, suggesting either a lack of genetic variation for this trait or the involvement of as yet unidentified factors underlying the differences in S. ratti fitness in rats and mice respectively. These results are discussed in relation to competing theoretical models of ecological specialization, host immunology and previous attempts to experimentally alter the host-specificity of parasitic nematodes

1. Six species of Amitermes, one from Panama and five from California, have been examined for protozoa. Certain protozoa have been found in many or all individuals examined of all species, but they are much less abundant than are flagellates in termites of the three xylophagous families below the Termitidae.

2. Trichomonas lighti n.sp. has been found in all species, but not in all individuals.

3. Chilomastix minuta n.sp. occurs in Amitermes beaumonti.

4. Embadomonas termitis n.sp. is described from Amitermes beaumonti.

5. Endamoeba beaumonti n.sp. has been observed in five host species. In its size and nuclear structure it resembles Endamoeba disparata of Mirotermes hispaniolae.

6. Nyctotherus silvestrianus n.sp. has been found in three of the Californian species. It resembles N. termitis and N. ovalis, but is larger than the former and smaller than the latter. A long filament is attached to the end of the cytopharynx. A thickened, V-shaped structure lies against the cyto-pharynx. Several deeply staining fibrils arise from this V.

7. A coccidian has been found in one individual of Amitermes minimus. This forms sixteen banana-formed merozoites and three sporoblasts as in Chagasella.

8. Micro-organisms other than protozoa are very abundant in the intestines of all six species of Amitermes, and they are similar in all.

Leishmania turanica n.sp., found infecting the desert rodent Rhombomys opimus in the southern territories of the USSR and the Mongolian People's Republic, is described. This parasite exists sympatrically with L. major and L. gerbilli in R. opimus and is the predominant species. A total of 284 isolates of L. turanica from R. opimus, 3 from naturally infected Phlebotomus andrejevi and 1 from P. papatasi were characterized and found to be clearly distinguishable on isoenzyme and nuclear DNA characteristics from all other Old World taxa of Leishmania.

The distribution of N. brasiliensis within the small intestine of rats, following a primary infection, is described.

The fourth stage larvae were found to inhabit the sections later parasitized by the adult population.

Both the larval and adult worms were found to be confined to the anterior half of the small intestine. This aggregation is believed to be under the influence of stimuli (factor(s) 1), present in the lumen of the host gut.

The adult worms also clumped within the already confined worm population. This behaviour is thought to be due to intrinsic behaviour patterns of the worm, sexual attraction and thigmokinesis.

Larval and adult worms were surgically transferred to points, at varying distance from the ‘normally’ inhabited region, in the small intestine of fresh recipient rats. The resulting migratory behaviour of the worms was studied.

The worm populations resulting from both the larval and the adult transfers showed the ability to recognize that they were in the wrong position of the small intestine and to be able to orientate, migrate and locate the ‘normal’ site.

The fine structure of the developing macrogamete of Eimeria maxima was studied from chicks killed at intervals from 138 to 147 h after inoculation. The macrogamete developed within a parasitophorous vacuole. Lying within this vacuole and extending for some distance around the periphery of the macrogamete were intravacuolar tubules, grouped in certain areas, and in some cases they were seen to make direct connexions with the cytoplasm of the parasite. During development, electron-pale vesicles were pinched off externally from the surface of the macrogamete. There appeared to be 2 forms of wall-forming bodies of the Type I during development, one form being less osmiophilic than the other. Other organelles present, such as wall-forming bodies of Type II, granular endoplasmic reticulum, mitochondria, canaliculi, lipid inclusions and intravacuolar folds, were similar in structure to those of other Eimeria species.

An isometric transducer system has been used to record spontaneous motor activity in Diclidophora merlangi in vitro. Motility took the form of either continuous irregular contractions or bursts of activity with intermittent quiescent periods. Maximal activity was recorded from specimens at 5–8 °C in artificial sea water (ASW). Decerebration induced a period of enhanced motility which subsided within 1 h. Water turbulence elicited large, rapid contractions of the longitudinal body musculature which did not habituate or fatigue. Induced water turbulence at frequencies higher than 3/min inhibited spontaneous movements, resulting in a progressive reduction in contraction amplitude and, eventually (30/min), the abolition of any response to water movement. The neurotoxin tetrodotoxin failed to modify worm activity. Excitatory responses from both intact worms and strip preparations were obtained with 5-hydroxytryptamine, dopamine and noradrenaline, but aminergic antagonists failed to block their effects. Atropine stimulated contractility, whereas the effects of acetylcholine or carbachol were inconsistent. Nicotine increased muscle tone but the effect was unaltered by atropine, mecamylamine or d–tubocurarine. Muscarine, hemicholinium, neostigmine and eserine were without apparent effect.

During the course of a field study of coccidiosis in turkeys, Eimeria oocysts were found which had much smaller dimensions than any previously recorded isolate from the turkey. These oocysts were purified by single oocyst infection of a turkey. The first oocysts (mean dimensions 16.15 ×14.75 μm) were recovered 103 h later. Inoculation of between 0·5 and 2.5 ×105 oocysts of this isolate caused severe effects on body weight gain. Cross-immunity studies showed the parasite to be a strain of E. meleagrimitis. Electrophoretic analyses of two enzymes showed that the strain could be differentiated from another strain of E. meleagrimitis. (Weybridge strain B).

The results show that strain variation occurs within the species E. meleagrimitis and extreme caution should be used in identifying species of Eimeria from the turkey by their oocyst characters.

Ce trématode a été trouvé par Nicoll dans l'intestin du Totanus calidris L. dans la Grande-Brétagne.

The object of this paper is to suggest that there may be an unexpected connexion between parasites and the evolution of sex, using for illustration an unfamiliar type of parasite, the selfish chromosome. The major intellectual challenge of sexuality is to an environment which is continually getting worse. The elegant solution given by the Red Queen theory (Levin, 1975; Hamilton, 1980; Bell, 1982; Bell & Maynard Smith, 1988) is that the relevant aspect of the environment is provided by antagonists—pathogens, predators and competitors—which, because they can respond adaptively so as to negate any improvement that has been made, provide a constant stimulus for continued evolution. Sexuality and recombination are favoured because some of the new combinations of genes which they create are resistant to the current population of antagonists. In other respects, sex and recombination are probably highly disadvantageous: outcrossed sex is expensive because it halves the rate of transmission of genes, while recombination breaks up successful combinations of genes. It is only in certain circumstances that the necessity for continual counter-adaptation will overcome these disadvantages: in particular, the damage (reduction in fitness) caused by an antagonist must be substantial, and the amount of damage must depend on a genetic interaction between the antagonistic species. These requirements are often satisfied by host—parasite systems, where both the ecological and genetic interactions between the antagonists may be very severe and highly specific (see reviews by Day, 1974 and Burdon, 1987). It is possible, therefore, that sex and recombination are maintained in natural populations largely through the dynamics of the coevolution of hosts and their parasites. This is certainly compatible with the major ecological patterns shown by sexual systems, with outcrossed sex being more common in the sea than in freshwater, more common at low than at high latitudes, and generally more common in stable, complex, climax environments where interactions between species are expected to be more frequent and intense (Bell, 1982). However, there is as yet no evidence which conclusively supports a direct causal link between the incidence of parasitism and the rate of recombination. In particular, it has never been demonstrated that a particular parasite has the effect of eliciting, directly or indirectly, a greater rate of genetic recombination in its host. We suggest that such a parasite exists; both the parasite and its effects are well known, but have never been interpreted in the context of the evolution of recombination through host—parasite coevolution. It is in many respects a rather unusual parasite. We shall argue that B-chromosomes represent highly evolved parasitic DNA, transmitted through the germ line and often eliciting greater rates of recombination in the host genome.

Many Neglected Tropical Diseases (NTDs) have recently been subject of increased focus, particularly with relation to high-throughput screening (HTS) initiatives. These vital endeavours largely rely of two approaches, in vitro target-directed screening using biochemical assays or cell-based screening which takes no account of the target or targets being hit. Despite their successes both of these approaches have limitations; for example, the production of soluble protein and a lack of cellular context or the problems and expense of parasite cell culture. In addition, both can be challenging to miniaturize for ultra (u)HTS and expensive to utilize. Yeast-based systems offer a cost-effective approach to study and screen protein targets in a direct-directed manner within a eukaryotic cellular context. In this review, we examine the utility and limitations of yeast cell-based, target-directed screening. In particular we focus on the currently under-explored possibility of using such formats in uHTS screening campaigns for NTDs.