Comparison of infection success,development and swim bladder pathogenicity of two congeneric Anguillicola species in experimentally infected Anguilla anguilla and A. japonica

Two closely related parasites, Anguillicola crassus and Anguillicola novaezelandiae, originally parasitizing swim bladders of the Japanese eel (Anguilla japonica and the Short-finned eel (Anguilla australis), respectively, were used for analyzing the infection success of each parasite species on either long-known, recently acquired or new definitive host species and the associated effects on the eels’ swim bladders. On that account, European eels (Anguilla anguilla) and Japanese eels were experimentally infected with both Anguillicola species in the laboratory. Susceptibility of the two eel species to both parasite species was determined by analyses of infection data. Subsequently, histopathological effects of the nematodes on the hosts’ swim bladders were characterized according to already established indices. The present study revealed significant differences between the four different host-parasite systems regarding recovery rates, infrapopulations, and damage levels. Both nematode species achieved significantly lower recovery rates in Japanese eels than in European eels, since the examined swim bladders of Japanese eels contained a high amount of dead encapsulated larvae, whereas those of European eels contained only living nematodes. Encapsulation of larvae in Japanese eels was associated with a distinct thickening of the swim bladder walls. The swim bladders of uninfected Japanese eels turned out to be generally thicker than those of European eels. Infection with both Anguillicola species resulted in a further thickening process of the swim bladder walls in Japanese eels, whereas those of European eels showed only minor changes. The two established classification systems turned out to be inapplicable, since the measurements and the macroscopic evaluations of the swim bladders of the two infected eel species did not entirely correspond to the underlying criteria.     

Ultrastructural study of a tetratrichomonad isolated from pig fecal samples

The migratory response of peripheral blood granulocytes and monocytes from the European eel Anguilla anguilla (L.) to infective larvae of the swimbladder nematode Anguillicola crassus Kuwahara, Niimi and Hagaki, 1974 was examined by means of light microscopical histology and with an in vitro assay using a modified Boyden chamber. Histological examination of experimentally infected eels revealed that, already 8 days postinfection, an infiltration of inflammatory cells around L₃ of A. crassus in the swimbladder tissue can occur. In the Boyden chamber, in presence of infective larvae of A. crassus (L₃), neutrophil granulocytes and monocytes showed a higher migration activity than in the absence of L₃. In conclusion, infection of European eels with A. crassus leads to an activation of the defence cells resulting in an increased migration activity compared to uninfected eels.     

Natural Anguillicola novaezelandiae infection—is there seasonality in New Zealand?

Knowledge of natural Anguillicola infections of Short-finned eels (Anguilla australis) in New Zealand is very limited. So far, no study contains data on all life cycle stages of Anguillicola novaezelandiae in naturally infected eels. In order to study the frequency of A. novaezelandiae in New Zealand Short-finned eels, we examined eels of the North and the South Island for the presence of the swim bladder parasite. The results show that A. novaezelandiae is a common parasite of the Short-finned eel. The parasite was present in both regions. Eels from both localities show differences in their infection status with respect to prevalence, abundance and intensity. While eels of the South Island were only infected with larval stages, adult and preadult stages could be detected in eels of the North Island. Nevertheless, infrapopulations at both sites were dominated by larval parasite stages. This unique composition of infrapopulations has never been described for any Anguillicola species before and suggests a seasonal occurrence as a possible reason. Export of live eels should be handled cautiously to prevent the spread of A. novaezelandiae throughout other eel populations.     

Interaction between the severity of the infection by the nematode <Emphasis Type="Italic">Anguillicola crassus</Emphasis> and the tolerance to hypoxia in the European eel <Emphasis Type="Italic">Anguilla anguilla</Emphasis>

An experiment was conducted to test the effect of the infection by the swimbladder nematode Anguillicola crassus on the survival of the European eel Anguilla anguilla when exposed to hypoxic conditions. Forty-four wild caught and naturally infected eels were placed in an aquarium filled with water from the fishing site (Vaccarès Lagoon, French Mediterranean coast). In this confined environment (271), under decreasing oxygen resources, the first eel death occurred after 45 h (O₂ = 0.98 mg l⁻¹) and the last one after 96 h (O₂ = 0.48 mg l⁻¹). After dissection and parasite examination, analyses revealed significant negative correlations between the time to death and various parameters of parasite pressure (e.g., number of lumen worms, parasite mass, health state of the infected organ). It was shown that the severity of damage to the swimbladder rather than the count of living parasites was the major contributing factor in explaining the variation in time to death. These semi-experimental data demonstrating an increased eel mortality rate under severe oxygen stress are discussed for their relevance under field conditions, especially during hot summer months.     

The Subcellular Location of Ovalbumin in Plasmodium berghei Blood Stages Influences the Magnitude of T-Cell Responses

Model antigens are frequently introduced into pathogens to study determinants that influence T-cell responses to infections. To address whether an antigen''s subcellular location influences the nature and magnitude of antigen-specific T-cell responses, we generated Plasmodium berghei parasites expressing the model antigen ovalbumin (OVA) either in the parasite cytoplasm or on the parasitophorous vacuole membrane (PVM). For cytosolic expression, OVA alone or conjugated to mCherry was expressed from a strong constitutive promoter (OVA_hsp70 or OVA::mCherry_hsp70); for PVM expression, OVA was fused to HEP17/EXP1 (OVA::Hep17_hep17). Unexpectedly, OVA expression in OVA_hsp70 parasites was very low, but when OVA was fused to mCherry (OVA::mCherry_hsp70), it was highly expressed. OVA expression in OVA::Hep17_hep17 parasites was strong but significantly less than that in OVA::mCherry_hsp70 parasites. These transgenic parasites were used to examine the effects of antigen subcellular location and expression level on the development of T-cell responses during blood-stage infections. While all OVA-expressing parasites induced activation and proliferation of OVA-specific CD8⁺ T cells (OT-I) and CD4⁺ T cells (OT-II), the level of activation varied: OVA::Hep17_hep17 parasites induced significantly stronger splenic and intracerebral OT-I and OT-II responses than those of OVA::mCherry_hsp70 parasites, but OVA::mCherry_hsp70 parasites promoted stronger OT-I and OT-II responses than those of OVA_hsp70 parasites. Despite lower OVA expression levels, OVA::Hep17_hep17 parasites induced stronger T-cell responses than those of OVA::mCherry_hsp70 parasites. These results indicate that unconjugated cytosolic OVA is not stably expressed in Plasmodium parasites and, importantly, that its cellular location and expression level influence both the induction and magnitude of parasite-specific T-cell responses. These parasites represent useful tools for studying the development and function of antigen-specific T-cell responses during malaria infection.     

Influence of site, season, silvering stage, and length on the parasites of the European eel Anguilla anguilla in two Mediterranean coastal lagoons of the island of Corsica, France using indicator species method

The parasites of 425 European eels, Anguilla anguilla, were studied between 2009 and 2012 in two Mediterranean coastal lagoons of the island of Corsica, France. An indicator value (IndVal) method was used for analysis, which combines measures of fidelity and specificity. Because of its resilience to detect changes in abundance, IndVal is an effective ecological bioindicator. The IndVal method demonstrated that site, season, silvering stage, and length could influence the occurrence of parasite species in European eel. A randomization test identified ten parasite species as having a significant indicator value for site (lagoons differed principally in salinity: oligohaline to polyhaline for the Biguglia lagoon and polyhaline to euhaline for the Urbino lagoon; the digeneans Bucephalus anguillae and Lecithochirium musculus, the cestodes Bothriocephalus claviceps, Proteocephalus macrocephalus, and larvae of Myzophyllobothrium sp., the nematodes Anguillicoloides crassus, and encysted larvae of Contracaecum sp., the acanthocephalan Acanthocephaloides incrassatus, the monogenean Pseudodactyogyrus anguillae, and the copepod Ergasilus gibbus); one parasite species for the spring season (the acanthocephalan A. incrassatus); six parasite species for silvering stage (yellow, pre-silver, silver; the trematodes B. anguillae and Deropristis inflata, encysted larvae of the nematode Contracaecum sp., the acanthocephalan A. incrassatus, the monogenean P. anguillae, and the copepod E. gibbus); and three parasite species for some of the five length classes (the cestode P. macrocephalus, encysted larvae of the nematode Contracaecum sp., and the monogenean P. anguillae). Data for species composition and infection levels should help to improve the management of parasitism in the populations of European eels.     

Gross and microscopic pathological changes associated with parasitic infection in European eel (Anguilla anguilla, Linnaeus 1758)

The gross pathological and histopathological changes associated with parasitic infection in the European eel (Anguilla anguilla Linnaeus, 1758) was investigated. A total of 65 eels collected from three sampling localities in Eastern Delta, Egypt were examined over the period of January–May 2008. The fish were subjected to standard procedures for parasitological and pathological examinations. Overall, 22 (33.8%) of the 65 fish examined were found to have parasitic infections. The eels harbored a total of six parasite species; among them, the nematode Anguillicoloides crassus was the most prevalent species (10.7%), followed by the Monogenea Pseudodactylogyrus anguillae (7.7%) and Dactylogyrus species (6.1%), the ciliate Trichodinella epizootica (4.6%), the Myxozoa Myxidium giardi (3.1%), and the cestode Proteocephalus macrocephalus (1.5%). Affected fish showed varying levels of tissue damage and pathological alterations including mild to severe degenerative, necrotic, and inflammatory changes in the affected organs.     

From yellow to silver: Transforming cranial morphology in European eel (Anguilla anguilla)

The European eel (Anguilla anguilla) has been extensively studied, especially because of its highly specialized migratory behaviour associated with substantial phenotypic transformations. During this migration, one of those transformations the eel undergoes is from yellow to silver eel, a process known as silvering. Although the cranial morphology during the earlier glass, elver and yellow eel stages are well studied, little is known about actual morphological changes during the transformation process from the yellow to the silver eel stage. Yet, literature suggests drastic changes in musculoskeletal anatomy. Here, we investigated the cranial musculoskeletal morphology of 11 male European eels at different stages during silvering, resulting both from natural and artificial maturation. Using 3D-reconstructed µCT data of the head, the skull and cranial muscles associated with jaw closing and respiration were studied. Eye size was used as a proxy for the silvering stage. Size-adjusted jaw muscle volumes increased during silvering, although insignificantly. Accordingly, a near-significant increase in bite force was observed. Respiratory muscles size did increase significantly during silvering, however. Considering the eel's long migration, which often includes deep and thus potentially oxygen-poor environments, having a better performing respiratory system may facilitate efficient migration. Both overall skull dimensions and specifically orbit size increased with eye index, suggesting they play a role in accommodating the enlarging eyes during silvering. Finally, artificially matured eels had a wider and taller skull, as well as larger jaw muscles than wild silver eels. This could be caused (a) by different conditions experienced during the yellow eel stage, which are maintained in the silver eel stage, (b) by side effects of hormonal injections or (c) be part of the maturation process as artificially induced silver eels had a higher eye index than the wild silver eels.     

Trypanosoma carassii hsp70 increases expression of inflammatory cytokines and chemokines in macrophages of the goldfish (Carassius auratus L.)

We report on the cloning and characterization of Trypanosoma carassii 70 KDa heat shock protein (hsp70). T. carassii hsp70 was secreted/excreted into culture medium in vitro and was recognized by sera from infected fish. Recombinant hsp70 (rhsp70) activated goldfish macrophages and stimulated the production of pro-inflammatory cytokines including interferon gamma (IFNγ), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, (IL)-12 and chemokines CCL-1 and CXCL-8 (IL-8). T. carassii hsp70-induced cytokine expression was abrogated by pronase treatment of macrophages confirming the existence of receptor(s) on goldfish macrophage surface that recognize parasite molecule. Parasite hsp70 also up-regulated the expression inducible nitric oxide synthase (iNOS) isoforms A and B and induced a strong nitric oxide response of goldfish macrophages.     

Parasite communities in eels of the Island of Reunion (Indian Ocean): a lesson in parasite introduction

Eel populations from the small rivers on the Island of Reunion (French Overseas Department in the Indian Ocean) were investigated with respect to the occurrence and abundance of helminths during the autumn of 2005. The native species Anguilla marmorata (n = 80), Anguilla bicolor (n = 23), and Anguilla mossambica (n = 15) were studied. Six species of helminths were identified, four of them having a definitely nonnative status. Furthermore, unidentified intra-intestinal juvenile cestodes and extra-intestinal encapsulated anisakid nematode larvae were present in a few eels. We found that the invasive swim bladder nematode Anguillicoloides (Anguillicola) crassus had been introduced into the island. Six specimens were collected, four from A. marmorata, one from A. bicolor and one from A. mossambica. The maximum intensity of infection was two worms. The other helminths also showed a low abundance. These species were the monogenean gill worms Pseudodactylogyrus anguillae and Pseudodactylogyrus bini and the intestinal parasites Bothriocephalus claviceps (Cestodes), Paraquimperia africana (Nematodes), and the acanthocephalan Acanthocephalus reunionensis Warner, Sasal, and Taraschewski, 2007. The latter species, found as intra-intestinal immatures, is thought to utilize amphibians as required hosts; its status, introduced or native, could not be determined. P. africana was described from A. mossambica in South Africa and has not been recorded outside Africa. The other species are known from populations of European and American eels. However, A. crassus and the two Pseudodactylogyrus species originate from East Asia, where they are indigenous parasites of Anguilla japonica. Both an assignment test based on seven specific microsatellite loci and subsequent sequencing of mitochondrial haplotypes of a partial fragment of cytochrome c oxidase 1 strongly suggest that the A. crassus may originated around the Baltic Sea. According to the results presented here, populations of the indigenous eel species from Reunion can be considered to harbor extremely isolationist alien parasite communities. Our findings support the hypothesis that during the present time of global biological change, invasion by a nonnative species into a target island is more likely to reflect the political affiliation of the colonized environment and the pathways of trade and tourism than geographic proximity between donor and recipient areas or other natural circumstances.     

expression of members of the heat-shock protein 70 family in the exoerythrocytic stages ofPlasmodium berghei andPlasmodium falciparum

Exoerythrocytic stages ofPlasmodium berghei cultured in HepG2-A16 hepatoma cells and those ofP. falciparum in human hepatocytes transplanted under the kidney capsule of CB-17/ICr scid/scid mice were used to evaluate expression of heat-shock-related stress proteins. Although undetectable in the sporozoites, the expression of proteins similar in sequence of a heat-shock protein of 70 kDa and a glucose-regulated protein of 78 kDa was markedly induced in the hepatic stages of malaria parasites. Expression of these proteins in the exoerythrocytic stages of the malaria parasite warrants a systematic evaluation of their potential role in eliciting cellular immune responses directed against infected hepatocytes.     

Temporal and spatial changes in the composition and structure of helminth component communities in European eels Anguilla anguilla in an Adriatic coastal lagoon and some freshwaters in Italy

The composition and diversity of the helminth component communities in eels Anguilla anguilla were determined in three separate localities in Italy: an Adriatic coastal lagoon, Comacchio and two freshwater localities, the River Po and the Lake Piediluco. Data from Comacchio lagoon were analysed over 15 years to determine whether community composition and diversity changed significantly overtime. The community was species rich (nine species, all marine except Proteocephalus macrocephalus) and was dominated by a suite of digeneans: Deropristis inflata, Helicometra fasciata, Lecithochirium musculus and Bucephalus anguillae. The community showed little change in composition over the period, but the relative abundance and dominance of the species did alter. By contrast, the component communities in the freshwater localities were species poor and the dominant species were freshwater acanthocephalans, Pomphorhyncus laevis in River Po and Acanthocephalus rhinensis in Lake Piediluco. The helminth community of Lake Piediluco with five species was richer than that of the River Po with only three species, but was poorer than that of Comacchio lagoons. Similarity indices between samples from Comacchio were high; between the lagoon and the freshwater localities and between the two freshwater localities, similarity indices were very low. Helminth component community structure in coastal lagoons was comparable across Europe. The helminth community in the River Po was similar to those in the River Tiber and other European rivers whilst that in Lake Piediluco was similar to that in other European lakes. Levels of the pathogenic Anguillicoloides crassus in swim bladders were consistently lower in prevalence and abundance in the coastal lagoons than in freshwater localities. This suggests that this parasite may have little impact on migrating eels if they are indeed primarily of marine origin and so it may be of little importance in the recent decline of eel populations throughout Europe.     

Novel DNA virus isolated from samples showing endothelial cell necrosis in the Japanese eel, Anguilla japonica

Economic loss due to viral endothelial cell necrosis of eel (VECNE) of Anguilla japonica is a serious problem for the cultured Japanese eel market. However, the viral genome responsible for VECNE is unknown. We recently developed a rapid determination system for viral nucleic acid sequences (RDV) to determine viral genome sequences. In this study, viral DNA fragments were obtained using RDV, and approximately 15-kbp circular full genome sequences were determined using a next-generation sequencing system, overlapping PCR, and Southern blot analysis. One open reading frame (ORF) was homologous to the large T-antigen of polyomavirus; other ORFs have no homology with any nucleic or amino acid sequences of polyomavirus. Therefore, as this DNA virus might comprise a novel virus family, we provisionally named it Japanese eel endothelial cells-infecting virus (JEECV). JEECV was detected in both naturally and experimentally infected eels, suggesting that JEECV potentially causes VECNE.     

Relationship between butyrylcholinesterase activity and liver injury in mice acute infected with Toxoplasma gondii

This study aimed to investigate the butyrylcholinesterase (BChE) activity in mice experimentally infected with Toxoplasma gondii during the acute phase. Twenty mice were divided in two groups with 10 animals each: group A was composed of uninfected mice while group B was formed by rodents infected with T. gondii. Five days after infection, blood was collected and serum separated, and fragments of liver and brain were obtained. In serum and liver homogenate was noted a significant reduction (P < 0.05) in BChE activity in infected mice when compared with uninfected ones. In serum was observed an increase in the activity of alanine aminotransferase and urea, associated with reduction in alkaline phosphatase activity and in the levels of total protein and albumin. Histologically, there were foci of necrosis and parasites in the forms of tachyzoites and cysts, with bradyzoites in liver samples of infected animals. Based on these results, we conclude that toxoplasmosis reduces BChE activity in mice, and this alteration is probably related to the liver damage caused by the parasitism. Therefore, this enzymatic alteration can directly contribute to the pathogenesis of the disease.     

Confirmation of the hosts involved in the life cycle of an acanthocephalan parasite of Anguilla anguilla (L.) from Lake Piediluco and its effect on the reproductive potential of its amphipod intermediate host

A total of 37 European eels, Anguilla anguilla, collected from Lake Piediluco, Central Italy, and measuring 35 to 75.5 cm in total length (mean±1 SD, 56.41?±?10.89 cm) were examined, and their acanthocephalan infections assessed. Thirty-two (86.49%) eels were infected with Acanthocephalus rhinensis (mean±1 SD, 67.38?±?65.16; range, 1-350), a species that, purportedly, can be discriminated on the basis of a characteristic band of orange-brown pigmentation encircling the anterior end of the trunk. This feature, however, was not seen on any of the A. rhinensis specimens that were removed, either attached to the gut wall or free within the gut lumen, from infected eels. Approximately 40% of the eels were coinfected with the dracunculid swimbladder nematode Anguillicoloides crassus, while a single eel was also coinfected with eight specimens of a second acanthocephalan, Dentitruncus truttae. From the stomachs of two eels, 109 intact and partially digested specimens of amphipod Echinogammarus tibaldii (Pinkster & Stock 1970) were recovered, 16 (14.6%) of these were infected with one to two cystacanths of A. rhinensis per host. From a sample of 850 E. tibaldii taken from the peripheral lakeside vegetation, 102 (12%; sex ratio, 1:1) gammarids were infected with one to two A. rhinensis cystacanths. Unparasitised ovigerous female E. tibaldii specimens had significantly higher numbers of eggs in their brood pouches compared with their infected counterparts (t-test, P?相似文献   

Dimorphism throughout the European eel's life cycle: are ontogenetic changes in head shape related to dietary differences?

A well‐known link exists between an organism's ecology and morphology. In the European eel, a dimorphic head has been linked to differences in feeding ecology, with broad‐headed eels consuming harder prey items than narrow‐headed ones. Consequently, we hypothesized that broad‐heads should exhibit a cranial musculoskeletal system that increases bite force and facilitates the consumption of harder prey. Using 3D‐reconstructions and a bite model, we tested this hypothesis in two life stages: the sub‐adult yellow eel stage and its predecessor, the elver eel stage. This allowed us to test whether broad‐ and narrow‐headed phenotypes show similar trait differences in both life stages and whether the dimorphism becomes more pronounced during ontogeny. We show that broad‐headed eels in both stages have larger jaw muscles and a taller coronoid, which are associated with higher bite forces. This increased bite force together with the elongated upper and lower jaws in broad‐headed eels can also improve grip during spinning behavior, which is used to manipulate hard prey. Head shape variation in European eel is therefore associated with musculoskeletal variation that can be linked to feeding ecology. However, although differences in muscle volume become more pronounced during ontogeny, this was not the case for skeletal features.     

Intra-population variation in behavior modification by the acanthocephalan Acanthocephalus dirus: are differences mediated by host condition?

The acanthocephalan parasite Acanthocephalus dirus infects the freshwater isopod Caecidotea intermedius as an intermediate host before completing its life cycle in a fish. Male C. intermedius infected by A. dirus parasites are less likely to engage in mating behavior than uninfected males but there is a significant intra-population variation in the occurrence of this behavioral change. Previous studies on uninfected isopods have shown that glycogen content is a predictor of male mating behavior and we examined whether the intra-population variation in the mating behavior of infected male C. intermedius could be explained by this relationship. A field-based behavioral experiment was used to quantify intra-population variation in male mating behavior, which showed that 50 % of infected males were responsive to females and 50 % were not responsive. Biochemical analysis of responsive and non-responsive males revealed that glycogen content was a predictor of the mating behavior for uninfected males but was not a predictor of mating behavior for infected males. For infected males, parasite intensity was a predictor of mating behavior. Males that contained more A. dirus parasites were less likely to undergo modification of mating behavior. We propose that the intra-population variation in the mating behavior of infected C. intermedius identified in nature was not mediated by host condition.