Numerical response of a helminth community in the course of a multi-annual abundance cycle of the water vole (Arvicola terrestris).

The impact of parasitism on population dynamics is determined in part by the numerical responses of parasites during population fluctuations of their hosts. Vole populations fluctuate in multi-annual cycles allowing such responses to be studied over successive phases of population growth, abundance and decline. We investigate how a helminth community (5 nematode and 7 cestode species) evolved over a full 6-year Water Vole (Arvicola terrestris) population cycle. Brillouin and individual parasite species richness (IPSR) indices were used to measure the numerical response of the parasite community. We report a correlation between levels of parasite intensity and vole population cycle phases. Both indices were consistently higher during pre-decline and decline phases for male and female voles alike. The numerical response of the parasite community suggests that populations may be regulated by parasitism and that studies of this mechanism should allow both for the cyclic or non-cyclic character of the host populations and for the response of the broadest possible set of the local parasite community.

Histomorphological study of the preputial and clitoral glands in BALB/c mice with experimental Taenia crassiceps infections.

Male preputial and female clitoral glands of mice undergo development that depends on the level of hormones in the animal. Experimental infection with Taenia crassiceps cysticerci results in significant physiological modifications in the host. Here, we investigated the histomorphological alterations induced by the parasite in these pheromonal glands. Preputial and clitoral glands were recovered from mice at 15, 35, 50, and 70 days postinfection (DPI). The glands were examined macroscopically and microscopically after histological preparation. Male preputial glands show a marked atrophy 35 days after infection. This atrophy is the result of a disorganization of the acinus tissue structure. During the course of infection, the basal, intermediate, and mature acinar cell layers are reduced, and finally, at 70 DPI, the gland includes only the duct system and fibrotic structures. In contrast, females are not affected by the infection because no modifications were observed in the morphology or histology of the clitoral glands. A probable cause for such a divergence between infected male and female mice might be related to a sex steroid imbalance as described during T. crassiceps infection.

Plasmodium chabaudi chabaudi chronic malaria and pathologies of the urogenital tract in male and female BALB/c mice.

BALB/c mice of both sexes were infected with a non-virulent strain of Plasmodium chabaudi chabaudi and any pathologies occurring in the urogenital tract and its accessory glands were investigated. Organs and tracts were removed from infected and control mice at 15, 40 and 100 days post-injection, weighed and processed for macroscopical and histological analyses. The relative weights of preputial, clitoral glands and testes were modified in infected mice during the 40 days following infection. The preputial glands show a marked hypotrophy at 15 days post-infection. The bladders of half of the infected female mice and a few infected male mice displayed a conspicuous haemoglobinuria and frequent interstitial cystitis that worsened throughout the experiment. Also, several chronic inflammatory reactions were detected in the prostates, preputial and clitoral glands up to 100 days post-infection. A probable cause of such a divergence in the characteristics of the infection and in the nature of the pathologies identified in infected male and female mice is the interaction between the infection and the immune and endocrine systems of the host. The cause of the pathologies and their consequences on the host condition are discussed.

Asexual multiplication of larval parasitic worms: a predictor of adult life-history traits in Taeniidae?

The hypothesis that asexual multiplication inside the intermediate host and adult life-history traits within the final host are independent is tested among Cestoda. Using phylogenetic relationships among the Cestoda species, we can show that asexual multiplication appears to have been lost and recovered several times throughout Taeniidae evolution; this allows a comparison of the adult life-history traits of species with and without asexual multiplication at the larval stage. The adult trait considered is the size of the parasite, since numerous life-history traits, such as fecundity and longevity, are correlated with size. If adult traits are independent of whether the larval stage reproduced asexually or not, we expect no difference in the adult size of the proliferative (i.e. with asexual multiplication) and non-proliferative species. The results are inconsistent with this hypothesis. In contrast, species with asexual multiplication in the intermediate host seem to have smaller adult size, reflecting a trade-off. We propose that ecological factors involving intraspecific and interspecific competition in the final host might be responsible for this trade-off. The role of these parameters in the evolution of life-history traits and more precisely in the acquisition of asexual multiplication is investigated with comparative analysis and discussed in the context of host-parasite interactions.

Behavioural dominance and Taenia crassiceps parasitism in BALB/c male mice.

Behavioural dominance relationships in mouse populations are based upon fighting and antagonistic behaviour. Social rank is affected by the physiological states present in the mice. Experimental infection by Taenia crassiceps cysticerci induced physiological disorders and disrupted the dominant-subordinate status. Infected male mice showed strong perturbations in territorial behaviour and aggressiveness. Infected dominant male mice did not show a significant reversal of dominance order compared to uninfected mice. In addition, during confrontation between naive infected and healthy mice, infected animals more often assumed a subordinate status than healthy ones. The effects of the infection by T. crassiceps were more likely to prevent adult male mice from becoming behaviourally dominant than to reverse existing dominance relationships. The results are discussed on the basis of the parasite manipulation hypothesis and host optimal foraging and decision-making theories.

Characterization of cDNA encoding a L37a ribosomal protein from Taenia crassiceps and its potential use in phylogenetic reconstructions.

In this study, we characterized for the first time the complete sequence of a L37a cDNA from a cestode specie: Taenia crassiceps. A phylogenetic analysis of L37a ribosomal proteins from distant animal species is presented and the potential use of such proteins in molecule-based phylogeny is discussed.

Parasite-altered host behavior in the face of a predator: manipulation or not?

Parasitologists have generally accepted the idea that parasite-induced alterations in host behavior increase the chance for parasite survival and transmission or ensure the completion of its life cycle. The aim of the present study was to investigate modifications in the behavior of Taenia crassiceps-infected BALB/c mice in the face of a predator. The experiments showed modifications in the response of infected mice in comparison with uninfected controls on exposure to a predator final host. However, different studies lead us to suggest that the observed modifications are likely to be a secondary effect of the impact of the parasite on host physiology and immunity that favors its development and proliferation.

Microhabitat distribution and coexistence of Microcotylidae (Monogenea) on the gills of the striped mullet Mugil cephalus: chance or competition?

The two monogenean species Metamicrocotyla cephalus and Microcotyle mugilis have specific microhabitats on the gills of Mugil cephalus on which they may or may not coexist. M. cephalus is found in sector 1 of the posterior hemibranch of arch I. M. mugilis is found on the filaments of sector 5 of the posterior hemibranch of arch I. The coexistence of these two monogenean species on the same fish does not induce a change in their respective distribution, except for a marked preference of M. cephalus for the left side and of M. mugilis for the right. This 15-month-long study shows that when the two species of monogeneans coexist the infection prevalence and intensity are higher than in cases of monospecific infection. The specific character of the microhabitat, the apparent absence of interspecific competition, and the existence of positive species interactions are discussed.

Echinococcus multilocularis protoscoleces and hepatic cell activity in vitro.

E. multilocularis protoscoleces were co-cultured with hepatic cells in the presence of IAR 20 or BALB/c 3T3 cells. Hepatocyte activity was determined by assaying transferrin and albumin secretion in culture media. The level of these 2 plasma proteins is higher in hepatic/BALB/c 3T3 co-culture medium. In the presence of parasites, the transferrin level is unchanged while the production of albumin is stimulated during the first 48 h. Our results suggest that the albumin production could be attributed to a complex cellular cooperation between hepatocytes and activated Kupffer cells as previously observed in the acute inflammatory reaction.

Echinococcus multilocularis: in vitro interactions between protoscolices and Kupffer cells.

Echinococcus multilocularis protoscolices collected from experimentally infected jirds were incubated for 2 weeks in rat hepatic cell cultures cocultivated with or without feeder cells (BALB/c 3T3 and IAR 20). Scanning and transmission electron microscopy studies were performed during the course of the culture period. Kupffer cells (Kc) were seen adhering to the anterior and posterior ends of the protoscolices. Some protoscolices were fixed to the cell monolayer by a cluster of Kc adhering to the posterior end of the parasite. These cells were phagocytosing the glycocalyx and the electron-dense distal end of the microthrix of the protoscolex tegument. An alteration in the superficial tegumental cytoplasm with extensive mitochondrial damage was also noted. The properties expressed by Kc against protoscolices in vitro might also be relevant for proliferation of metacestodes in vivo.

Synthesis of propargylic alcohols and biological effects on Echinococcus multilocularis metacestodes.

This study describes the synthesis of propargylic alcohols derived from isatin and their biochemical and morphological effects on Echinococcus multilocularis metacestodes in Meriones unguiculatus. Propargylic alcohols decreased the alkaline phosphatase and the lactate dehydrogenase activities of the metacestode selectively. The most effective compound, 1b, decreased the lactate dehydrogenase enzymatic activity, and the glucose concentration in the parasite increased, whereas the glycogen content was partially decreased. Furthermore, the ultrastructure study revealed several damages. The host-parasite relationships are very important in the intrahepatic cestodes as shown by the biochemical side effects observed in the host's liver during the treatment. An in vitro enzymatic study was performed with alcohols 1b and 1c.

Developmental changes of Echinococcus multilocularis metacestodes revealed by tegumental ultrastructure and lectin-binding sites.

Ultrastructural investigations (SEM, TEM) combined with lectin-binding analysis, have revealed concurrent modifications in tegumentary structure and surface glycoconjugates during the establishment and differentiation of Echinococcus multilocularis metacestodes in jirds. The laminated layer, which is amorphous and rich in polysaccharides when initially secreted by the young cyst, takes on a different appearance and has a different glycoconjugate composition according to whether the cyst becomes fertile or sterile. The laminated layer of fertile cysts transforms into a microfibrillar matrix, the protein content of which may increase while sugar content decreases during protoscolex differentiation. Independently of this structure, brood capsules, from which arise protoscoleces, are formed by invagination of the cyst tegument. The intense secretion of glycoconjugates from the brood capsule wall during invagination may serve to interact with host factors passing through the laminated layer. The combined use of ultrastructural study and lectin labelling has allowed the demonstration of an ultrastructural and biochemical gradient of differentiation of the protoscolex. Seven stages of differentiation have been described. The possibility that the excreted-secreted tegumentary glycoconjugates, revealed by lectin labelling during protoscolex differentiation, might be the gradual biochemical expression of one or several stimuli implicated in the phenomenon of protoscolex maturation, is discussed.

In vivo cultivation of Echinococcus multilocularis protoscoleces in micropore chambers.

Micropore chambers containing unevaginated protoscoleces of E. multilocularis were implanted into the peritoneal cavity of AKR mice. Transformation from protoscoleces to fertile multivesicular cysts was obtained after 210 days. Ultrastructural observations of these morphological transformations indicate that a phase of histogenesis follows a phase of dedifferentiation. This morphogenetic process raises the question of the origin of new cell populations. The results reveal the potential role of protoscoleces in secondary echinococcosis and the value of this experimental model for further studies on the larval development.

A model for studying isolation mechanisms in parasite populations: the genus Lepeophtheirus (Copepoda, Caligidae).

In the Mediterranean, the parasitic copepod Lepeophtheirus thompsoni Baird, 1850 specifically infests turbot (Psetta maxima L., 1758), whereas L. europaensis Zeddam, Berrebi, Renaud, Raibaut, and Gabrion, 1988 infests brill (Scophthalmus rhombus L., 1758) and flounder (Platichthys flesus L., 1758). Experimental infestation of turbot by copepods from each of the three fish species showed an absence of any physiological incompatibility preventing natural development of the two parasite species, at least on one host species, i.e., the turbot. Moreover, interspecific hybrids were obtained experimentally, which implies that 1) there is no strict genetic barrier between the two species and 2) the natural prezygotic isolation results from a choice of the most favorable habitat. We discuss the origin and possible consequences of the presence, in the Mediterranean, of L. europaensis on brill and flounder, two hosts separated by their taxonomic status and ecobiology.

Selective distribution of sugars on the tegumental surface of adult Bothriocephalus gregarius (Cestoda: Pseudophyllidea).

Whole specimens and histological and semi-thin sections of Bothriocephalus gregarius adults were exposed to lectins to identify carbohydrates present in the tegument and parenchyma. The sugars N-acetyl glucosamine, N-acetyl galactosamine, galactose, glucose (or mannose) and fucose were detected in the cestode using eight lectins: WGA (Wheat germ agglutinin), HPA (Helix pomatia agglutinin), SBA (Soy bean agglutinin), PHA (Phaseolus vulgaris agglutinin), RCA60 and RCA120 (Ricinus communis toxin and agglutinin), ConA (Concanavalin agglutinin) and UEA-I (Ulex europaeus agglutinin). Combined use of these methodological approaches (whole specimens, paraffin and semi-thin sections) revealed the presence of a gradient in the distribution of most of the sugars over the tegument, with the highest concentrations on the strobila (as shown by most of the lectins). Other sugars were specific for the scolex or strobila (as shown by UEA-I or HPA, respectively). The ultrastructural study showed that the distribution of glycoconjugates was associated with the presence of specific tegumental coats. The significance of this selective distribution and its relevance to cestode physiology and host-parasite relationships are discussed.

The fine structure of the embryonic envelopes before and after hatching in bothriocephalids: physiological and ecological significance.

The fine structure and differentiation of the embryonic envelopes before and after hatching in two bothriocephalid species of teleostean fish are described. Scanning electron microscopic observation of the egg surface confirmed the specific ornamentation of the capsule. Its thickness and topographic morphology reflect the physiocochemical characteristics of the environment. Beneath the eggshell, two syncytial layers were observed around the oncosphere. The outer envelope, visible during the preoncospheral phase, degenerated before the liberation of the coracidium. Glycoconjugate labelling confirmed the nutritive character of this layer. After hatching, the inner envelope was the only layer covering the oncosphere. The apical plasma membrane of the coracidial sheath bore cilia and numerous clublike microvilli. Beneath the cortical zone, the syncytial cytoplasm was vacuolated, resembling a honeycomb; the electron-dense zone, described from other pseudophyllideans, was not present. The basal membrane of the inner envelope, uniformly electron-dense, was assimilated by the oncospheral membrane. Comparison of the composition of the embryonic envelopes of pseudophyllideans with those of other tapeworm orders and trematodes revealed ontogenetic, structural and functional similarities between the embryotrophic layers. The physiological and ecological significance of the structure of the free-swimming coracidium is discussed. In this respect, the inner envelope around the oncosphere appears to be responsible for the inability of the larva to detect and invade the intermediate host.

Echinococcus multilocularis metacestodes: biochemical and ultrastructural investigations on the effect of isatin (2-3 indoline dione) in vivo.

A study on the biochemical and ultrastructural effects of isatin on Echinococcus multilocularis metacestodes in Meriones unguiculatus is reported. In the metacestode, after an 18 day treatment period, the alkaline phosphatase and lactate dehydrogenase activities decreased and the acid phosphatase activity increased. Glucose and glycogen stores declined significantly. Examinations of the ultrastructure revealed an increase of lysosomes and phagocytic reactions. The protoscolesces were damaged and an abnormal striated process appeared. The tegumental cytoplasmic syncytium remained unaltered. These observations suggest that isatin derivatives may have a role in the chemotherapy of infections caused by Echinococcus spp.

The effect of isatin on the Echinococcus granulosus cyst in an experimental host.

The effect of isatin on the E. granulosus cyst was studied. NMRI mice, which were infected with E. granulosus of sheep origin, were treated daily with isatin at a dose of 50 mg kg-1 for 18 days. Ultrastructural damage was observed in the treated cysts, including accumulation of lamellar stacks, electron-dense granules, autophagosomes and lipid vesicles. Moreover, a biochemical study showed an inhibition of alkaline phosphatase activity, with a decrease in carbohydrate storage and an increase in acid phosphatase activity. In spite of the short duration of the treatment, the results obtained allowed us to conclude that isatin acts on E. granulosus cysts. This activity appears as a process of degeneration linked to the alkaline phosphatase inhibitory effect of isatin.

Speciation of cestoda. Evidence for two sibling species in the complex Bothrimonus nylandicus (Schneider 1902) (Cestoda: Cyathocephalidea).

Using biochemical genetic methods, we have distinguished 2 sibling species in the complex Bothrimonus nylandicus (Schneider, 1902), which infest 2 congeneric species of sole (Solea lascaris and Solea impar) on European coasts (Atlantic and Mediterranean). Neither of the parasite species is specific for either of the sole species, but one of them is present all year round, whereas the other is absent in the autumn and winter and only appears in the spring, subsequently disappearing at the end of the summer. Only S. impar lives in the Mediterranean, and is equally infested by both cestodes, whereas both species occur in the Atlantic and each of them is preferentially infested by 1 species of cestode. The shortness of the adult stage of the parasite in the definitive host and the presence of 2 life-cycles associated with competition between the 2 hosts in the Atlantic could be responsible for the biological differences observed and for maintaining the sibling species in sympatry.