We tested the chemotactic responses of dauer juvenile stages (DJs) of the insect parasitic nematode Heterorhabditis bacteriophora to a variety of compounds that are known to be highly attractive or highly repellent to Caenorhabditis elegans. While H. bacteriophora DJs respond to alcohols and some aromatic compounds as well as to host metabolites such as uric acid and CO2, the most notable difference in the responses of these two nematodes is that H. bacteriophora DJs are unresponsive to a large number of compounds which C. elegans finds highly attractive. The latter compounds are typical by-products of bacterial metabolism and include aldehydes, esters, ketones and short-chain alcohols. While C. elegans finds long-chain alcohols (e.g. 1-heptanol and 1-octanol) repellent and short-chain alcohols highly attractive, H. bacteriophora DJs are strongly attracted to 1-heptanol, 1-octanol and 1-nonanol and find short-chain alcohols to be only slightly attractive. Parasitic-stage H. bacteriophora nematodes show a very weak chemotactic response to volatile molecules that DJs find highly attractive. Our results suggest that, associated with the adoption of a parasitic mode of life by Heterorhabditis, there was an adaptive change in chemotactic behaviour of the infective stages, resulting in a decreased sensitivity to volatile by-products of bacterial metabolism and an increased sensitivity towards long-chain alcohols and other insect-specific volatiles and possibly also to herbivore-induced plant volatiles.

Trypanosomes isolated from South American bats include the human pathogen Trypanosoma cruzi. Other Trypanosoma spp. that have been found exclusively in bats are not well characterized at the DNA sequence level and we have therefore used the SL RNA gene to differentiate and characterize kinetoplastids isolated from bats in South America. A Trypanosoma sp. isolated from bats in southern Brazil was compared with the geographically diverse isolates T. cruzi marinkellei, T. vespertilionis, and T. dionisii. Analysis of the SL RNA gene repeats revealed size and sequence variability among these bat trypanosomes. We have developed hybridization probes to separate these bat isolates and have analysed the DNA sequence data to estimate their relatedness. A new species, Trypanosoma desterrensis sp. n., is proposed, for which a 5S rRNA gene was also found within the SL RNA repeat.

TaqMan minor groove binder probes were evaluated as to their suitability for the real-time allelic discrimination of the β-tubulin codon 200 TTC/TAC single nucleotide polymorphism in cyathostomin species. Amplification of titrated cloned full-length β-tubulin cDNA revealed that the TaqMan minor groove binder PCR is capable of specifically detecting as few as 10 copies. Testing of DNA from single adult and larval stages of several different species of cyathostomin allowed reproducible genotyping of individual worms. Using the real-time PCR approach, the throughput of samples was considerably increased compared with conventional post-PCR readout procedure. Only 7·8% homozygous TAC L3 were found among 102 L3 which were genotyped from phenotypically BZ-resistant small strongyle populations. The percentages of the homozygous TTC and heterozygous TTC/TAC were 41·3% and 50·9%, respectively. This resulted in a total TAC-allele percentage of only 33·3%. These findings correspond to data obtained by genotyping of an experimentally selected BZ-resistant cyathostomin population. It is concluded that the β-tubulin codon 200 polymorphism is not the sole mechanism involved in the process of BZ resistance in cyathostomins.

In this study, we conducted phylogenetic analyses of nematode parasites within the genus Dictyocaulus (superfamily Trichostrongyloidea). Lungworms from cattle (Bos taurus), domestic sheep (Ovis aries), European fallow deer (Dama dama), moose (Alces alces), musk ox (Ovibos moschatus), red deer (Cervus elaphus), reindeer (Rangifer tarandus) and roe deer (Capreolus capreolus) were obtained and their small subunit ribosomal RNA (SSU) and internal transcribed spacer 2 (ITS2) sequences analysed. In the hosts examined we identified D. capreolus, D. eckerti, D. filaria and D. viviparus. However, in fallow deer we detected a taxon with unique SSU and ITS2 sequences. The phylogenetic position of this taxon based on the SSU sequences shows that it is a separate evolutionary lineage from the other recognized species of Dictyocaulus. Furthermore, the analysis of the ITS2 sequence data indicates that it is as genetically distinct as are the named species of Dictyocaulus. Therefore, either this taxon needs to be recognized as a new species, or D. capreolus, D. eckerti and D. viviparus need to be combined into a single species. Traditionally, the genus Dictyocaulus has been placed as a separate family within the superfamily Trichostrongyloidea. The present molecular phylogenetic analyses support the placement as a separate family, but the current data do not support the placement of the Dictyocaulidae within the Trichostrongyloidea without a reassessment of the placement of the superfamily Strongyloidea. While D. eckerti has been regarded as the one and only lungworm species of cervids, this study showed that 4 host species including 3 members of Cervidae (moose, reindeer, red deer) and 1 Bovidae (musk ox) were infected with this parasite. Host ranges of D. viviparus (cattle), D. filaria (sheep) and D. capreolus (moose and roe deer) were more restricted. No clear pattern of co-evolution between the dictyocaulid taxa and their bovid and cervid hosts could be determined.

An assessment is made of the repeatability of parasite community structure in space for a marine fish, and in space and time for a freshwater fish from south-eastern Mexico. The marine fish species was the red grouper, Epinephelus morio (collected from 9 localities), and the freshwater species was the cichlid, Cichlasoma urophthalmus (collected from 6 localities: including monthly at 2 localities for 1 year, and bimonthly at 1 locality in 1990 and 1999). Pairwise interspecific associations and analyses of nested patterns in the distributions of parasite species among hosts were used in both fish species, with comparisons over time made only with the cichlid. Positive interspecific associations, and nested patterns were noted in some localities for both fish species, and/or at some sampling times for the cichlid fish. However, non-random patterns in the structure of parasite communities in these 2 host species only were observed sporadically. When present, nestedness in both fish species was apparently linked with a positive association between total infection intensities and fish size. Additionally, adjacent localities were more likely to display similar parasite community structure than distant ones. This preliminary result suggests that distance between localities is an important determinant of predictability in parasite community structure.

The saliva of haematophagous insects has a series of pharmacological activities which may favour blood feeding. In the present study, an inhibitory effect on the complement system was observed in salivary extracts obtained from the phlebotomine sand flies Lutzomyia longipalpis and Lu. migonei. Saliva from Lu. longipalpis was capable of inhibiting both the classical and alternative pathways, while that from Lu. migonei acted only on the former. Other haematophagous insect species were screened for inhibition of the classical pathway. The triatomine bugs Panstrongylus megistus, Triatoma brasiliensis and Rhodnius prolixus were also able to inhibit the classical pathway whereas the mosquito Aedes aegyti and flea Ctenocephalides felis were not. The activity of Lu. longipalpis saliva on the classical pathway was partially characterized. The inhibitor is a protein of Mr 10000–30000 Da, which is very resistant to denaturation by heat. The inhibition of the complement system by phlebotomine sand flies may have a role in the transmission of Leishmania to the vertebrate hosts. The inhibitor molecule is thus a promising component of a vaccine to target salivary immunomodulators.

Howardula helenoschini sp. n. (Tylenchida: Allantonematidae) is described from 40 million year-old Baltic amber. The nematodes are parasitic in the body cavity of an adult fly of the genus Triphleba (Phoridae: Diptera). Characteristics of the fossil species are the large size of the pre-adult juveniles and a small number of progeny compared with extant Howardula parasites of flies. Reasons are given why these features suggest that this species was at an early stage of adapting, evolutionarily, to its insect host.

Thirty-one stocks of Trypanosoma cruzi, the agent of Chagas disease, representative of the genetic variability of the 2 principal lineages, that subdivide T. cruzi, were selected on the basis of previous multilocus enzyme electrophoresis analysis using 21 loci. Analyses were performed with lower numbers of loci to explore the impact of the number of loci on the robustness of the phylogenies obtained, and to identify the loci that have more impact on the phylogeny. Analyses were performed with numerical (UPGMA) and cladistical (Wagner parsimony analysis) methods for all sets of loci. Robustness of the phylogenies obtained was estimated by bootstrap analysis. Low numbers of randomly selected loci (6) were sufficient to demonstrate genetic heterogeneity among the stocks studied. However, they were unable to give reliable phylogenetic information. A higher number of randomly selected loci (15 and more) were required to reach this goal. All loci did not convey equivalent information. The more variable loci detected a greater genetic heterogeneity among the stocks, whereas the least variable loci were better for robust clustering. Finally, analysis was performed with only 5 and 9 loci bearing synapomorphic allozyme characters previously identified among larger samples of stocks. A set of 9 such loci was able to uncover both genetic heterogeneity among the stocks and to build robust phylogenies. It can therefore be recommended as a minimum set of isoenzyme loci that bring maximal information for all studies aiming to explore the phylogenetic diversity of a new set of T. cruzi stocks and for any preliminary genetic typing. Moreover, our results show that bootstrap analysis, like any statistics, is highly dependent upon the information available and that absolute bootstrap figures should be cautiously interpreted.

An area close to the Qinghai-Tibet plateau region and subject to intensive deforestation contains a large focus of human alveolar echinococcosis while sporadic human cases occur in the Doubs region of eastern France. The current review analyses and compares epidemiological and ecological results obtained in both regions. Analysis of rodent species assemblages within quantified rural landscapes in central China and eastern France shows a significant association between host species for the pathogenic helminth Echinococcus multilocularis, with prevalences of human alveolar echinococcosis and with land area under shrubland or grassland. This suggests that at the regional scale landscape can affect human disease distribution through interaction with small mammal communities and their population dynamics. Lidicker's ROMPA hypothesis helps to explain this association and provides a novel explanation of how landscape changes may result in increased risk of a rodent-borne zoonotic disease.

The toxicity of cadmium and zinc at concentrations ranging from 0·1 to 10 000 μg/l to the life-span of decaudised cercarial bodies (cercariae that have shed their tails) of Diplostomum spathaceum was investigated. The effects of metal exposure at 3 temperatures (12, 20, and 25 °C) and 3 levels of water hardness (distilled water, soft water and hard water) were studied. In general, under most experimental conditions increasing metal concentrations reduced the life-span of decaudised cercariae. Increasing water hardness and decreasing water temperature caused an increase in the life-span of both control and metal exposed decaudised cercariae. However, at certain isolated metal concentrations, associated with a specific level of water hardness and temperature, increased survival above controls occurred. Differences in the relative toxicity of cadmium and zinc were dependent on the environmental conditions of exposure. The decaudised cercarial life-span under metal exposure was found to be generally independent of the overall cercarial survival and tail loss in most experimental conditions. Prolonged exposure to cadmium and zinc caused changes in the decaudised cercarial life-span when compared to individuals decaudised during the initial 24 h exposure period to those which were decaudised during the final 24 h period of cercarial survival. The validity of studying the decaudised cercarial life-span as an indicator of ‘fitness’ of larvae to migrate through the tissues of the target fish host, in terms of glycogen utilization, was assessed for those cercariae decaudised during the initial 24 h exposure period only. A limited reduction in the decaudised cercarial life-span during this period compared to controls was recorded, which may possibly indicate a reduced penetration ‘fitness’ of cercariae exposed to cadmium and zinc. The importance and relevance of these findings to parasite migration and establishment in the fish host are discussed.

Chemotactic responses of Strongyloides stercoralis infective larvae (L3) to sodium chloride (NaCl) were investigated by recording larval tracks on a saline gradient in agarose. On agarose, larvae migrated randomly, whereas when placed at 0·01 M NaCl larvae moved to approximately 1·1 M NaCl where they turned, headed down the gradient and eventually remained circling at a favoured salinity (0·03–0·07 M). Conversely, when placed at 2·85 M NaCl, the L3 larvae moved unidirectionally to lower, more favoured salt concentrations. Here they circled, changing directions frequently while making ‘loop-like’ tracks. Larvae were immobilized within 5 min at salt concentrations exceeding 3 M NaCl. When placed at 0·01 M NaCl, 51·1%±26·9 migrated to 1·1 M NaCl after 2 min, and 80%±18·7 did so after 8 min, at an average velocity of 4·1±1·4 mm/min. Larvae (53·6%±21·6) were repelled from 2·85 M NaCl to lower concentrations after 2 min. After 8 min, 95%±11·1 were repelled, moving at an average velocity of 6·2±1·1 mm/min. Using this bioassay, the influence of neuronal control over chemotactic behaviour of S. stercoralis and other parasitic nematodes can be elucidated.

Since 2 morphological forms (fertilized and unfertilized) of egg can be produced by Ascaris, infected humans can release in their faeces fertilized eggs only (FEO), unfertilized eggs only (UEO) or both fertilized and unfertilized eggs (FUE) (designated herein as the 3 different egg profiles). Epidemiologically, fertilized eggs are of significance as they enable effective transmission of the parasite. This study, for the first time, characterizes the Ascaris egg profiles in human faeces in an endemic region of China, explores possible host- and parasite-factors related to these profiles, and discusses the biological and epidemiological implications of the findings. The 3 egg profiles were recorded throughout the study period of 2 years, and the overall percentages of people with FEO, FUE and UEO profiles were ~41–47%, 32–42% and 17–21%, respectively. The overall number of unfertilized eggs for the entire population accounted for ~6–9% of all eggs excreted. The different Ascaris egg profiles showed no correlation to host gender, but they did relate to age and worm burden of the host and to the sex ratio and developmental status of the parasite. While an annual universal anthelmintic treatment resulted in some fluctuation in the values of individual egg profiles, the general features of these profiles remained similar throughout the study period. The findings of this study should have significant implications for understanding transmission patterns of Ascaris and for the implementation of control measures against ascariasis in endemic regions.

The aim of this paper was to test the hypothesis that when the 2 species of Acanthocephalan Pomphorhynchus laevis and Acanthocephalus clavula are found concurrently within the intestine of brown trout under field conditions, they have the potential to interact negatively. Evidence has shown that Acanthocephala are more likely to exhibit negative interactions with their own and other species, under both field and experimental conditions. Furthermore, the likelihood of these interactions is increased in Ireland because of the absence of certain definitive hosts and the fact that concurrent infections by two or more species of Acanthocephala are more commonly observed in fish. Data collected from wild and stocked brown trout and from 2 lakes provided an opportunity to compare the 2 potentially interacting helminth species in their fundamental and realized niche and several pieces of convincing evidence are provided here to support the hypothesis. A significant negative association between the numbers of each species found in individual fish was reported and this was consistent for both wild and stocked trout. Furthermore, an analysis of the proportions of low, moderate and high intensity infections in single and concurrent infections revealed a significant reduction in increasing intensities in concurrent infections compared to single infections. Finally, strikingly different patterns of niche inhabitation were observed, particularly for P. laevis in the presence of A. clavula in wild trout. Results from the niche width analysis also support the observations on average position in single and concurrent infections. The niche width of P. laevis when it co-occurred with A. clavula decreased markedly in high intensity infections compared to low intensity infections.

In theory, the age at which maturation occurs in parasitic nematodes is inversely related to pre-maturational mortality rate, and cross-species data on mammalian nematodes are consistent with this prediction. Immunity is a major source of parasite mortality and parasites stand to gain sizeable fitness benefits through short-term adjustments of maturation time in response to variation in immune-mediated mortality. The effects of thymus-dependent immune responses on maturation in the nematode parasites Strongyloides ratti and Nippostrongylus brasiliensis were investigated using congenitally thymus-deficient (nude) rats. As compared with worms in normal rats, reproductive maturity of parasites (presence of eggs in utero) in nude rats occurred later in S. ratti but earlier in N. brasiliensis. Immune-mediated differences in maturation time were not associated with differences in worm length. Thymus-dependent immunity had no effect on pre-maturational mortality. Results are discussed in relation to theoretical expectations and possible explanations for the observed patterns in parasite maturation.

Risk factors for the transmission of Echinococcus multilocularis to humans operate at a range of spatial scales. Over a large area, such as China, regional scale risk is correlated with variation in climatic conditions because of its effect on the spatial distribution of landscapes that can support E. multilocularis transmission in wildlife hosts and the probability of egg survival. At a local scale of a few kilometres, or tens of kilometres, transmission risk is related to the spatial proximity of human populations and landscapes with active transmission. At the patch scale, when considering individual villages or households, human behavioural factors are important and for individuals genetic and immunological factors play a role. Satellite remote sensing can provide landscape information at a range of spatial scales and provide a spatial framework within which to examine transmission patterns. This paper reviews the application of remotely sensed data and spatial data analysis to develop a better understanding of disease transmission and shows how such data have been used to examine human alveolar echinocossosis infection patterns, at a range of spatial scales, in an endemic area in central China.

The population dynamics of Toxocara canis in pigs, and their immune response to a primary and a challenge infection, were studied by parasitological, haematological and serological parameters. Seventy pigs were divided into 4 groups; 35 pigs received a primary infection (group A), 15 pigs received both a primary and a challenge infection (group B), 15 pigs received the challenge infection only (group C), and 5 pigs served as helminth-free controls (group NC). A dose of 50000 eggs was administered for the primary infection (day 0) and a dose of 10000 eggs was given for the challenge infection (day 28). On days 7, 14, 21 and 28 p.i., 5 pigs of group A, and on days 35, 42 and 49 p.i., 5 pigs from each of groups A, B and C were necropsied. Numbers of recovered larvae varied widely among the 5 pigs of each group on all days of necropsy. Toxocara canis larvae were recovered predominantly from the lungs; migration of larvae to other organs or tissues from the lungs was restricted. In group A, the larval burden in the lungs peaked on day 14 p.i., and the larval densities decreased significantly over time. Thereafter, the majority of larvae were recovered from the lungs until the end of the experiment (day 49 p.i.). A few larvae were found in the muscles and brain until day 42 p.i., and 2 larvae were found in the eyes of 2 pigs on day 35 p.i. There was little evidence of protective immunity to a challenge infection in this experiment. The eosinophil levels tended not to increase in pigs receiving a challenge infection, in contrast to the challenge control pigs. The fact that T. canis larvae migrate and persist in the tissues of pigs for more than 1 month suggests a zoonotic risk in infected pigs. The relevance of these data to the population biology and immunology of porcine and human toxocarosis is discussed.

Cystic echinococcosis, caused by the larval stage of Echinococcus granulosus, is a global public health problem. Whilst in a few localities, such as New Zealand, the parasite has been effectively controlled or even eradicated, in most endemic regions it remains a persistent problem. In some areas, such as the former Soviet Union, the disease incidence in humans has increased rapidly in recent years. It is important to have an understanding of the transmission dynamics, both between dogs and domestic livestock where the parasite maintains itself and from dogs to people. It is from this knowledge that effective control measures can be devised to reduce the prevalence of the parasite in animals and hence reduce the incidence of human disease. Mathematical models to describe the transmission of the parasite and the effects of different control strategies were first proposed over twenty years ago. Since then further information has been acquired, new technology has been developed and better computing technology has become available. In this review, we summarise these developments and put together a theoretical framework on the interpretation of surveillance information, how this affects transmission and how this information can be exploited to develop new intervention strategies for the control of the parasite. In particular, the parasite remains a persistent or re-emerging problem in countries of low economic output where resources for an intensive control programme, that has been successful in rich countries, are not available. By understanding of the transmission biology, including mathematical modelling, alternative and cost-effective means of control can be developed.

Analyses of small subunit ribosomal RNA gene sequences of representatives of major taxa of Monopisthocotylea were performed to identify the sister group of Gyrodactylus. Nuclear ribosomal DNA sequencesNucleotide sequence data reported in this paper are available in the GenBank, DDBJ and EMBL databases under the accession numbers AJ566375-79, AJ566768, AJ567670-74, AJ581657. from the complete internal transcribed spacer (ITS) region were used to infer phylogeny of 37 Gyrodactylus species and Gyrodactyloides bychowskii, Macrogyrodactylus polypteri and Gyrdicotylus gallieni, using maximum likelihood, parsimony and Bayesian inference. The genus Gyrodactylus appeared to be a monophyletic group in all analyses, based on the present data set. Within the genus, there were 3 major groups recognized by high bootstrap values and posterior probabilities. None of the 6 subgenera appeared to be monophyletic, and the most basal subgenus G. (Gyrodactylus) was paraphyletic. Characteristics of the excretory system of Gyrodactylus do not seem to be conservative enough to reveal subgenera within Gyrodactylus and we suggest abandoning existing subgenera as indicators of phylogeny. The grouping of species based on the morphology of the ventral bar and marginal hooks seems to have sufficient power to infer relationships between the Gyrodactylus species.

A total of 35 pigs aged 15 weeks old, and 21 pigs aged 8 weeks old were divided into 7 groups. Groups 1 and 2 were uninfected and challenge control groups, respectively. Groups 3 and 4 were infected weekly with 6 increasing doses of Ascaris suum eggs, and group 4 was additionally treated with pyrantel. Groups 5, 6, and 7 were immunized weekly with the 14, 42, or 97 kDa fractions from adult worms, respectively. Animals of groups 2–7 were challenged with 10 000 A. suum eggs 7 days after the last infection/immunization. Serum was sampled weekly and specific IgG1, IgG2, and IgM responses were measured. Pigs of groups 5, 6, and 7 showed high IgG1 and IgG2 responses especially against adult worms antigens, while infected groups had high IgG1 and IgM responses, especially against larva. The IgG1 responses were negatively correlated to the numbers of larvae in the lungs, and positively associated with the liver white spot numbers. There was a positive correlation between IgG2 and the numbers of white spots and lung larvae, while IgM was negatively correlated with these parasitological measures. These findings are discussed and it is suggested that acquired resistance against A. suum larvae is correlated with the induction of IgG1 and IgM, and not with IgG2, and that future vaccination protocols may focus on inducing the Th2 activity.

Following a brief introduction of recent advances in molecular and immunological technology for detection of persons and animals infected with Echinococcus multilocularis and an overview of the current situation of alveolar echinococcosis (AE) in Japan, perspectives on control options are discussed with reference to different epidemiological situations. AE is considered the most serious parasitic zoonosis in temperate and arctic regions of the northern hemisphere. The number of human cases differs drastically among regions. While high numbers of patients are apparently associated with high E. multilocularis prevalence in domestic dogs, e.g. in parts of Alaska and western China, the number of cases is moderate or low in areas where the parasite is mainly transmitted by wild canid species (e.g. in central Europe or temperate North America). However, the severity of the disease, the absence of curative treatment for most cases, the high cost of long-term chemotherapy and the anxiety caused for the population in highly endemic areas call for the development of preventive strategies even in regions where human AE is rare. Furthermore, in view of (1) drastically increasing numbers and infection rates of foxes involved in transmission of E. multilocularis, and (2) increasingly close contact between humans and foxes e.g. in Europe and Japan, there is considerable concern that AE incidences may in future increase in these regions. Control options depend on a variety of factors including the species of canid principally responsible for transmission and the socio-economic situation in the region. Where domestic dogs (stray or owned) are the principal hosts for E. multilocularis, control options can include those applicable to E. granulosus, i.e. reduction of the number of stray dogs, registration and regular preventive chemotherapy of owned dogs, and information campaigns for the population promoting low-risk behaviour for man and dogs. Where E. multilocularis is mainly transmitted by wild canids, the situation is far more difficult with preventive strategies still being in trial stage. Integrated control measures could include prevention information campaigns, restricting access of pet animals (dogs and cats) to rodents, chemotherapy of foxes on local or regional scales, and strategies to minimize contacts between people and foxes.