The attachment of Eimeria tenella to its target cells as an obligatory intracellular pathogen is essential for the development of disease. Previous reports have established that other intracellular protozoa parasites have either fibronectin, an adhesion protein, or fibronectin receptors, both of which are involved in the interaction with the host cells. In this current research, studies have been undertaken to visualize a surface component that may be involved in the attachment of E. tenella to host cells. For this purpose, monoclonal antibodies, both anti-chicken and anti-human fibronectin, and also anti-chicken integrin were used. Our results show a fibronectin-like molecule with an apparent molecular weight of 110 kDa in mature schizonts and microgametes. Staining with serum directed against chicken integrin revealed immunoreactivity within mature schizonts. Both the fibronectin-like molecule and the integrin may play an important role in the parasite stage-cell interaction and the promotion of parasite uptake.

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.

Recombinant Schistosoma mansoni elastase was expressed in Escherichia coli and an antiserum raised against the re-combinant protein was used to investigate stage-specific control of elastase in the parasite, and to determine whether the enzyme could form the basis of a strategy to prevent larval invasion of the host. Results showed that the expression of elastase is developmentally regulated, even if the basal promoter activity does not seem to be stage specific. The analysis of mRNA expression showed the presence of elastase transcript in adult worms although we could not detect the protein at this stage, suggesting that S. mansoni employs a form of translational control. The measurement of elastase levels in supernatants of culture schistosomula combined with the localization of elastase in cercariae invading mouse skin showed that the enzyme is heavily released during penetration. Finally, we studied the cytotoxic activity of rat anti-elastase sera, and the analysis of the isotypic profile suggested that IgG2a anti-elastase may be responsible for the cytotoxic effect.

Surveys on Trichinella parasites in domestic and sylvatic animals collected in France, Italy, and in the Extremadura region of Spain showed that the distribution of Trichinella spiralis and Trichinella britovi is influenced by both environmental and human behaviour factors. In France, both Trichinella species are prevalent in the fox population from mountain areas and natural parks but are infrequent in wild boars (< 0·001%). In Italy, only T. britovi is present in sylvatic animals (foxes, wolves, and mustelids) living 500 m above sea level. This species is rare in wild boars ( < 0·001%) in that area. Sylvatic trichinellosis is found in only 24% and 34% of French and Italian territory, respectively, while lowland areas may generally be considered Trichinella-free, because the domestic cycle is absent. The ecology of T. spiralis and T. britovi in the Extremadura shows a different picture from that observed in France and Italy because of the presence of both domestic and sylvatic cycles. The domestic cycle not only allows the maintenance of T. spiralis in the domestic environment, but it also has a great impact on the prevalence in wild boar populations. It does not influence the prevalence in vulpine populations. These data suggest (1) that domestic trichinellosis occurs only in rural areas of Western Europe in association with traditional swine-rearing practices, but not in industrialized pig farms; (2) that sylvatic trichinellosis occurs only in natural habitats which, in Western Europe, are widespread in mountain areas; (3) that the fox is the primary reservoir in the sylvatic cycle, where the parasite is maintained in a closed circuit and (4) that among sylvatic animals T. spiralis is present at lower altitude than is T. britovi.

Trichinella spiralis larvae infect and develop within skeletal muscle cells causing major changes to their mechanical properties. The aim of this investigation was to determine the effects of T. spiralis on the power output and fatigue resistance of the mammalian diaphragm under conditions simulating in vivo operation and to relate these to respiratory performance. Infection with T. spiralis leads to major reductions in mechanical stress, work, power output and fatigue resistance. These changes are associated with the number of larvae present in the muscle and the duration of infection. However, the initial decline in mechanical performance occurs during the onset of infection when there are few larvae observed within the muscle cells, indicating that T. spiralis may affect the properties of muscle before encapsulation. This may correspond to the host's inflammatory response and the effects of larval excretory/secretory products. The decline in mechanical performance will have a profound effect on respiration both at rest and during exertion. This must influence the behaviour of the host and increase its chance of capture by predators, which is likely to benefit the parasite by facilitating its transmission.

A longitudinal investigation on natural populations of Ascaris in humans and pigs and an investigation of soil contamination with Ascaris eggs were carried out from June 1993 to June 1994 in 2 villages, Manhu area, Xinjian County, Jiangxi Province, China. Results from these studies indicate that although human ascariasis is endemic there is significant fluctuation in both prevalence and the mean number of eggs/g faeces (epg) of the communities. Fluctuation of age-stratified prevalence and mean epg was detected in children but not in most adult groups. Most cases of human ascariasis were judged to involve low intensities of infection and a typical overdispersion distribution pattern was observed through the year. It was estimated that during the year, nearly half of the eggs discharged in the environment came from infections in children aged between 2 and 15 years which accounted for about 30% of the total population. Soil in and around houses and in vegetable gardens was found to be contaminated by Ascaris eggs and this situation remained relatively stable throughout the year. Monthly developmental rate of Ascaris eggs in soil was detected and the results suggest that the fluctuation in prevalence observed during the year should be directly attributed to the effect of seasonality of egg development. Features of Ascaris infection in pigs were found to be similar to those in humans except for a lower mean intensity of infection. The possibility of cross-infection of Ascaris between human and pig hosts is discussed.

The free radical nitric oxide (NO), which is synthesized by nitric oxide synthase (NOS), has recently been discovered to function as a neuronal messenger. The presence of NOS was detected in the nervous system of adult Hymenolepis diminuta with NADPH-diaphorase (NADPH-d) histochemistry. The NADPH-d histochemical reaction is regarded as a selective marker for NOS in neuronal tissue. NADPH-d staining was observed in nerve fibres in the main and minor nerve cords and the transverse ring commissures, and in cell bodies in the brain commissure, along the main nerve cords, in the suckers and the rostellar sac. NADPH-d staining was also observed in the wall of the internal seminal vesicle and the genital atrium. The pattern of NADPH-d staining was compared with that of the 5-HT immunoreactive nervous elements. The NADPH-d staining reaction and the 5-HT immunoreactivity occur in separate sets of neurons. This is the first time the NADPH-d reaction has been demonstrated in the nervous system of a flatworm, indicating that NOS is present and that NO can be produced at this level of evolution.

On the basis of known DNA sequences of Dirofilaria repens and D. immitis we designed specific primers for the amplification by Polymerase Chain Reaction (PCR) of the DNA from the two species. The PCR-based identification was found to be unambiguous and allowed specific diagnosis of microfilariae in blood samples, of developing larvae in the mosquito vector and of immature adults in bioptic material, overcoming the serious constraints of the morphological separation of these filarial parasites at the pre-adult stages. The technique was found to be very sensitive and applicable to samples stored either dry or in various preservation media, with the exception of formalin. The reliable identification of D. repens and D. immitis from bioptic material is expected to greatly enhance the chances of detecting human infections and to further clarify the role of the two parasites as pathogens of man. The possibility of routine identification of developing larvae in the vector will substantially improve the perspectives for epidemiological investigations, particularly in Southern European regions, such as Italy, where the two nematode species are largely sympatric.

Steinernema carpocapsae (Rhabditida: Steinernematidae) host recognition behaviour was assessed and compared with 2 measures of host suitability. Previous research showed that S. carpocapsae infective juveniles respond to host cues in a hierarchical order, with attraction to Galleria mellonella volatiles being stimulated by contact with G. mellonella cuticle. We measured host recognition behaviour by calculating the percentage response of S. carpocapsae infective juveniles to volatiles produced by G. mellonella last instars after the nematodes were exposed to the cuticle of 11 candidate arthropod hosts and 2 control surfaces. Host suitability was measured by nematode-induced mortality to candidate hosts at 2 nematode doses and the level of reproduction supported by each host. The highest recognition response was scored for Agrotis ipsilon (Lepidoptera: Noctuidae). This insect also incurred nearly 100% mortality due to nematode infection and supported the highest level of reproduction. Non-insect arthropods tested (Chilopoda and Isopoda) stimulated no behavioural response and were not susceptible to nematode infection. Other insect species elicited intermediate levels of the recognition response. There were significant correlations between behavioural response and nematode-induced mortality at the lower dose. The level of reproduction supported by the candidate hosts was also correlated with S. carpocapsae behavioural response.

A cDNA clone encoding a full length putative collagen has been isolated in a screen of a mixed stage Globodera pallida expression library. Comparison of the deduced amino acid sequence of this molecule with other collagens suggests it is a cuticular collagen and a member of the col-8 subfamily of collagen genes. Northern blots show the gene is expressed specifically in gravid, adult females of the parasite as compared to second (invasive) stage juveniles and virgin females. Preliminary immunocytochemical studies indicate this collagen is present in areas other than the cuticle; these findings and the potential functional role of this collagen are discussed.

Secretions of plant-parasitic nematodes which are released into plant tissue may play critical roles in plant-nematode interactions. The identification and characterization of these molecules are of fundamental importance and may help to facilitate the development of novel strategies to interfere with nematode infection of plants and thereby decrease nematode-induced damage to crops. An antibody-based approach was used to isolate molecules present on the nematode surface and in nematode secretions. Monoclonal antibodies (MAbs) were produced to secretions and to whole Heterodera avenue 2nd-stage juveniles; several of these MAbs recognized molecules present in nematode secretions produced in vitro. Three of these molecules have been partly characterized in H. avenae, Globodera rostochiensis, G. pallida and Meloidogyne incognita. A MAb reacting with the surfaces of these nematodes recognized antigens of different molecular weight in each of the species tested. This difference in antigenicity might be related to specific functions in these nematodes. Preliminary results show that this antibody also localized the antigen in root cells surrounding the feeding site induced by M. incognita in Arabidopsis thaliana.