The bacterial flora of the intestine plays an important role in the virulence caused by Entamoeba histolytica. Cysteine proteinase (CP), an amoebic virulence factor, plays a major role in host cell destruction. The mechanism of increased virulence following bacterial co-association is not understood. We studied CP of E. histolytica HM1:IMSS which was co-associated with Escherichia coli K12 strain pre-incubated with GalNAc or CP specific inhibitor E 64. Co-association of E. histolytica with bacteria enhanced CP activity 3·6-fold as assessed by azocasein assay and substrate gel electrophoresis showed bands at molecular weights of 28, 35 and 56 kDa. Northern and Western blot analysis showed increase in ehcp2 and ehcp5 gene expression. Trophozoites co-associated with E. coli showed greater cytotoxicity of BHK cells by a 51Cr release assay than trophozoites that had not been co-associated; this enhancement was abolished by E-64 treatment. The killing of BHK 21 targets by E. histolytica was characterized by DNA laddering which was not inhibited with E-64. GalNAc pre-incubation of trophozoites reduced cytotoxicity and DNA laddering, while E. coli co-associated E. histolytica showed smearing with faint laddering of BHK implicating both necrosis and apoptosis. Hence, bacterial co-association increases CP activity and CP gene expression and contributes to the necrosis of the target cell.

The neuromuscular systems of parasitic helminths are targets that are particularly amenable for anthelmintics. In this study, we describe a GABAergic neurotransmission in adult Schistosoma mansoni, the trematode responsible for high levels of morbidity in people living in developing countries. GABA immunoreactivity (GABA-IR) was detected in nerve cells and fibres of the cerebral ganglia and longitudinal nerve cords and the nerve plexuses ramifying throughout the parenchyma of male adult worms. In addition, strong GABA-IR was also found associated with the oral and ventral suckers as well as in testes indicating a role for GABA in fixation to the host vascular wall and spermatogenesis. The capacity to synthesize GABA from glutamate was confirmed by measurement of a glutamate decarboxylase (GAD) activity. Supporting these data, a single band with an apparent molecular weight of about 67 kDa was detected using an antibody raised against mammalian GAD. In vivo studies revealed that picrotoxin, a non-competitive antagonist of the GABAA receptor, produced a modification of the motility and locomotory behaviour of adult worms, suggesting that GABAergic signalling pathway may play a physiological role in the motonervous system of S. mansoni and could be considered as a potential target for the development of new drugs.

This study reports the distribution of parasite genotypes for the trypanosome Crithidia bombi across individual units (the colonies) in host populations of a social insect, the bumble bee Bombus terrestris. A number of microsatellite primers were developed and several of them were found to be polymorphic in our samples. Furthermore, a simple algorithm was used to identify the likely multi-locus genotypes present in multiply infected host individuals. The results demonstrated a remarkably high degree of genetic diversity among infections. A first sample from 1997 could only use a low resolution with 2 loci and showed a total of 11 different genotypes of C. bombi from 12 colonies. The sample from 2000 was analysed at 6 polymorphic loci and contained data from 8 colonies that were infected by 27 different C. bombi genotypes. Roughly 16% of all individual bees but half of all colonies (2000 sample) were infected with more than 1 genotype. The infections in the different colonies were also genetically distinct from each other, and the parasite population as a whole was in linkage disequilibrium and deviated from Hardy–Weinberg expectations. The highly structured and genetically diversified population of C. bombi is likely to result from strong genotypic host–parasite interactions.

This paper examines the efficiency of acquired resistance in protecting the fish host, rainbow trout (Oncorhynchus mykiss), against the trematode parasite Diplostomum spathaceum, and the hypothesis that fish recognize areas where infective stages are aggregated and show avoidance behaviour. We found that when fish with a low level of infection were held in restricted cages in natural conditions they became infected and developed cataracts as a result of this infection. This suggests that acquired resistance is insufficient in protecting fish against the parasite or the deleterious effects of infection in conditions where fish could not avoid the parasite. Behavioural experiments in the laboratory showed that fish reacted to the parasite cercariae by avoiding the infection source, which decreased the rate of parasite establishment. We conclude that by using a combination of behavioural avoidance and physiological resistance, fish could defend against the parasite more effectively.

The ultrastructure of the developing excretory system of Prosorhynchoides gracilescens and Prosorhynchus squamatus cercariae is described. The development pattern was similar in both species. In early embryos the two main collecting tubes were composed of a layer of cells which were wrapped around the lumen. Later, the tubes fused and the excretory epithelium of the fusion zone and that of the lateral caudal ducts became a syncytium. The collecting tubes in the cercarial body retained their cellular organization. As the tails grew, additional excretory pores were formed in the tail stem where thickened portions of the caudal duct epithelium contacted the surface tegument. Following this, the distal portions of the lateral caudal ducts lost contact with the primary excretory pores and progressively degenerated. Excretory atrium development started with differentiation of secretory active cytons peripheral to the fusion zone. These cells gave rise to cytoplasmic extensions that penetrated the fusion zone wall to eventually form a continuous cytoplasmic layer. This layer eventually replaced some of the fusion zone excretory epithelium and became the lining of the excretory atrium. The anterior end of the fusion zone differentiated into an excretory bladder and a short posterior portion gave rise to the caudal vesicle.

The anterior adhesive mechanism was studied for Merizocotyle icopae (Monogenea: Monocotylidae). Adult anterior apertures can open and close. In addition, duct endings terminating within the apertures are everted or retracted depending on the stage of attachment. Adhesive in adults is synthesized from all 3 secretory types (rod-shaped, small and large spheroidal bodies) found within anterior apertures. All exit together and undergo mixing to produce the adhesive matrix, a process that depletes duct contents. A greater number of ducts carrying rod-shaped bodies is depleted than ducts containing spheroidal bodies which changes the ratio of secretory types present on detachment. Detachment involves elongation of duct endings and secretion of additional matrix as the worm pulls away from the substrate. The change in secretory type ratio putatively modifies the properties of the secreted matrix enabling detachment. Only after detachment do ducts refill. During attachment, individual secretory bodies undergo morphological changes. The larval and adult adhesive matrix differs. Anterior adhesive in oncomiracidia does not show fibres with banding whereas banded fibres comprise a large part of adult adhesive. The data suggest that this is the result of adult spheroidal secretions modifying the way in which the adult adhesive matrix forms.

Allozyme variation within and among populations of 3 species of the genus Lecithochirium (Trematoda: Hemiuridae) was studied by starch gel electrophoresis. In total, 19 loci were analysed in 7 populations. The level of genetic variability was relatively high in all populations. The percentage of polymorphic loci (0·95 criterion) observed per population varied from 21·0% to 55·5%, and expected heterozygosity levels varied from 0·082 to 0·197. All populations showed significant heterozygote deficiencies. In Lecithochirium fusiforme most of the deviations from Hardy–Weinberg proportions were within the populations and this species showed moderate population structuring (FIS=0·486, FST=0·142, Nm=1·51) and accordingly low intraspecific genetic distances (D=0·003 to 0·027). A significant lack of heterozygotes for several polymorphic loci was revealed in Lecithochirium rufoviride and Lecithochirium musculus. The most probable cause of the population genetic subdivision in L. rufoviride is the presence of at least 1 cryptic species in the populations studied. Although the lowest percentage of fixed genetic differences was that between L. fusiforme and L. musculus, two different algorithms for the construction of evolutionary trees on a matrix of genetic distances confirmed that L. fusiforme and L. rufoviride are phenetically the most closely related species.

A nested (non-random) species composition was observed among the intestinal macroparasite communities of European eels, Anguilla anguilla. Nestedness was observed in 8 out of 10 component community samples from the rivers Thames and Test between April 2000 and October 2001. Parasite component communities consisted of mainly generalist and specialist, but also accidental species, and included acanthocephalans, cestodes, nematodes and digeneans. Nestedness was observed when component population size differed markedly between all or most parasite species, but not when the majority of species present was similarly abundant. Nestedness could not be explained in terms of host weight, log normal distribution of parasite species, or mean intensity of infection. It is proposed that nestedness occurred as a result of a sufficiently graded abundance between parasite species, which was established early in the year by colonization processes.

Here, the validity of the assumption of concerted evolution of ribosomal regions in larval and adult Cooperia oncophora was assessed. In each of 4 individuals of this parasitic nematode, at least 78% of the sequences comprised different ITS variantsNucleotide sequence data are available in the DDBJ/EMBL/GenBank databases under the Accession numbers AJ544390–AJ544465.. This implies that concerted evolution is not acting, which is corroborated by the scarcity of signatures of gene conversion and recombination. Mis-incorporation of nucleotides and illegitimate PCR-induced recombination turned out to be unlikely, and positions with substantial frequencies of alternative nucleotides corresponded to ambiguous positions in published ITS2 sequences of this and other Cooperia species based on direct sequencing. The ITS regions of each individual C. oncophora displayed a significant excess of unique mutations in agreement with expansion of the ribosomal gene family. Interesting corollaries of the inferred size changes of this gene family are genomic rearrangements that occur during larval development such as multiple rounds of endoduplication (in Rhabditidae), chromatin diminution (in Ascaris), and non-compensatory mutations on the secondary structure of the ITS2. It is yet unknown which process is important in trichostrongylids. Finally, although it can not be rigorously assessed in Cooperia, the ITS polymorphisms can readily be envisioned to affect phylogenetic reconstructions of closely related nematodes.

This paper reports a longitudinal study, conducted in 1989/90, of 1893 children aged 5 to 14 years in 9 poor urban areas of the city of Salvador (population 2·44 million), capital of Bahia State in northeast Brazil. Stool examinations were performed to measure nematode infection and reinfection 9 months after treatment, and an extensive questionnaire was applied to collect information on each child and on the conditions of the household. Comparison of areas with different levels of infrastructure showed the following trends as the level of community sanitation improved: clustering of cases by household became more significant, predisposition of individuals to reinfection and to heavy infection became more marked, and infections with different species were increasingly aggregated in the same individuals. These results suggest that sewerage and drainage can significantly reduce transmission of intestinal nematode infections in the public domain, but that other measures are required to control transmission within the household.

Homologues of c-fos and c-jun from total DNA of Taenia crassiceps and Taenia solium were cloned and sequenced. The amino acid alignment analysis revealed that c-fos DNAs from T. crassiceps and T. solium were highly homologous (96%), and both have high homology compared to several mammalian c-fos proteins (93% to mouse, 96% to rat and 86% to human). The c-jun protein alignment showed higher homology (T. crassiceps and T. solium have 98%), when compared with mouse, rat and human, being 92%, 98% and 93% respectively. RT–PCR amplification of the parasite's total RNA, showed that T. crassiceps expressed both AP-1 complex genes, while T. solium only expressed c-fos. Southern blot hybridization analysis confirmed the true origin of each amplified gene. AP-1 transcription gene expression is regulated by oestradiol in the same fashion as their mammalian counterparts only in T. crassiceps. To study if AP-1 genes are involved in a physiological function of the cyst, reproduction was studied in vitro. Oestradiol treatment stimulated reproduction in T. crassiceps but not in T. solium cysticerci. This is the first report of the detection and functionality of AP-1 transcription factor genes in any species of helminth parasite.

Gastrointestinal helminth infections remain a major constraint to livestock production globally. This study evaluated anthelmintic efficacy of 7 plants used as dewormers by farmers and pastoralists in Kenya. Thus 3 commercial anthelmintics and 7 plant preparations were tested in lambs infected with 5000 or 3000 L3 Haemonchus contortus in 4 experiments. In the first experiment, ivermectin, levamisole and albendazole were tested in 46 lambs. Seven plant preparations of Hagenia abyssinica, Olea europaea var. africana, Annona squamosa, Ananas comosus, Dodonea angustifolia, Hildebrandtia sepalosa and Azadirachta indica were tested in 151 lambs in 3 experiments. All 3 anthelminitics were highly effective in reducing faecal egg counts (FEC) and total worm counts (TWC) in lambs. Plant preparations had varying levels of crude proteins from 2·6% for O. europaea to 18·4% for A. indica. Compared with controls, no significant reductions in FEC were observed for any of the treated groups either 2 or 3 weeks post-treatment. Lambs treated with A. squamosa and A. comosus were slaughtered 4 weeks post-treatment. No significant differences were observed in mean TWC or number of eggs per female worm between treated animals and the controls. No significant improvements in weight gain were observed in treated lambs.