Swim bladder nematodes (Anguillicoloides crassus) disturb silvering in European eels (Anguilla anguilla)

The introduced parasite Anguillicoloides crassus is thought to play an important role in the decline of freshwater eel (Anguilla spp.) populations. These nematodes are known to negatively affect many fitness-related traits in eels. We used experimental infections to study the effect of A. crassus on the relative size or mass of organs, and the expression of functionally relevant genes (total of 12 parameters) that are involved in the silvering process of Anguilla anguilla. Our results showed that the liver mass, the hemoglobin α-chain, and androgen receptors α expression levels were significantly higher in infected eels, whereas the freshwater rod opsin expression level and the gut mass were significantly lower in infected eels. Our results suggested that infected eels were at a more advanced stage in the silvering process than uninfected counterparts of similar size. These results may be explained by 2 hypotheses. First, A. crassus could trigger physiological mechanisms involved in the silvering process as a side-effect of infection. Second, eels may adjust their life history traits in response to infection. The implications for eel migration and reproductive success may be either negative or positive, depending on whether the response to A. crassus infection results in an additional cost of the parasite or is due to the phenotypic plasticity of the host.     

Anal redness in European eels as an indicator of infection by the swimbladder nematode, Anguillicola crassus

Anal redness in European eels Anguilla anguilla is related to the prevalence and mean abundance of the swimbladder nematode Anguillicola crassus and may provide a simple, non-invasive diagnostic tool for A. crassus infection.     

The Giant Mottled Eel,Anguilla marmorata,Uses Blue-Shifted Rod Photoreceptors during Upstream Migration

Catadromous fishes migrate between ocean and freshwater during particular phases of their life cycle. The dramatic environmental changes shape their physiological features, e.g. visual sensitivity, olfactory ability, and salinity tolerance. Anguilla marmorata, a catadromous eel, migrates upstream on dark nights, following the lunar cycle. Such behavior may be correlated with ontogenetic changes in sensory systems. Therefore, this study was designed to identify changes in spectral sensitivity and opsin gene expression of A. marmorata during upstream migration. Microspectrophotometry analysis revealed that the tropical eel possesses a duplex retina with rod and cone photoreceptors. The λ_max of rod cells are 493, 489, and 489 nm in glass, yellow, and wild eels, while those of cone cells are 508, and 517 nm in yellow, and wild eels, respectively. Unlike European and American eels, Asian eels exhibited a blue-shifted pattern of rod photoreceptors during upstream migration. Quantitative gene expression analyses of four cloned opsin genes (Rh1f, Rh1d, Rh2, and SWS2) revealed that Rh1f expression is dominant at all three stages, while Rh1d is expressed only in older yellow eel. Furthermore, sequence comparison and protein modeling studies implied that a blue shift in Rh1d opsin may be induced by two known (N83, S292) and four putative (S124, V189, V286, I290) tuning sites adjacent to the retinal binding sites. Finally, expression of blue-shifted Rh1d opsin resulted in a spectral shift in rod photoreceptors. Our observations indicate that the giant mottled eel is color-blind, and its blue-shifted scotopic vision may influence its upstream migration behavior and habitat choice.     

Isolation of freshwater and deep-sea type opsin genes from the common Japanese conger

Two rod-like opsin genes, conf and cond , were isolated from the common Japanese conger Conger myriaster . The predicted proteins of conf and cond consisted of 354 and 350 amino acid (aa) residues respectively, and the aa identity was 87.4%. Phylogenetic analysis suggested that conf and cond are homologous to the freshwater type and the deep-sea type rod opsin genes of freshwater eels. RT-PCR analyses revealed that in both juveniles and adults cond is expressed in the retina and pineal complex, while conf is only expressed in the pineal complex at a low level.     

The swimbladder nematode Anguillicola crassus in the European eel Anguilla anguilla and the Japanese eel Anguilla japonica: differences in susceptibility and immunity between a recently colonized host and the original host

The swimbladder nematode Anguillicola crassus originates from Asia where it is a parasite of the Japanese eel Anguilla japonica. After its introduction to Europe about 25 years ago, the parasite spread rapidly within the indigenous populations of the European eel Anguilla anguilla and subsequently the prevalence and mean intensity appeared to stabilize. Under experimental and aquaculture conditions the na?ve new host appears to be more susceptible to A. crassus compared to the original host. Both eel species develop a immune response against A. crassus. The antibody response is well characterized for the European eel, but poorly characterized for the Japanese eel. It remains unclear if antibodies have any protective function against A. crassus. Encapsulation of larvae of A. crassus can be observed in naturally infected European eels. However, encapsulation of larvae following experimental infection has not been detected in European eels, but only in Japanese eels. Reinfection experiments and intraperitoneal injection of A. crassus homogenates failed to demonstrate the development of acquired immunity in European eels. Immunization with irradiated third stage larvae provided preliminary evidence for acquired immunity against A. crassus in the Japanese eel, but not in the European eel.     

First record of ostracods as natural intermediate hosts of Anguillicola crassus, a pathogenic swimbladder parasite of eels Anguilla spp

The ostracod Physocypria nipponica (Ostracoda: Candonidae) was found (prevalence 14.2%) to be the only intermediate host of the nematode Anguillicola crassus (Nematoda: Anguillicolidae), a pathogenic swimbladder parasite of eels, in a greenhouse-heated culture pond at Isshiki, Aichi Prefecture, Japan. Japanese eels Anguilla japonica from the same pond were found to be infected by adult A. crassus (prevalence 71.8%, intensity 1 to 6). This indicates that A. crassus could complete its life cycle under conditions of modern eel-culture technology where copepods were absent due to the unfavorable water quality for them, by utilizing ostracods as the intermediate host.     

Influence of Anguillicola crassus (Nematoda) and Ichthyophthirius multifiliis (Ciliophora) on swimming activity of European eel Anguilla anguilla

We investigated the swimming activity of 70 European eels Anguilla anguilla in relation to natural infection with 2 parasite species: the eel-specific swimbladder nematode Anguillicola crassus and the non-specific skin and gill protozoan Ichthyophthirius multifiliis. We measured how long individual eels exposed to a water current in a swimming channel with a steady-stream profile could withstand the water current. The parasites affected the swimming behaviour of eels in different ways. The maximum period of time the fish were able to swim against the current was not correlated with infection by A. crassus. In contrast, infection with I. multifiliis reduced the swimming time. The protozoan has a higher pathogenicity than the swimbladder nematode, at least in closed systems, where I. multifiliis is able to spread within a few days. Reduction in swimming capacity after infection with the ciliate averaged 47 % compared to capacity prior to infection. Thus, our results do not support the previously suggested strong negative relation between swimming activity of eels and intensity of A. crassus infection, at least in the short-term. However, there are indications in the literature that the pathological effects of A. crassus on the eel swimmbladder may involve a higher energy demand, possibly manifested in a prolonged spawning migration. As a result, eels heavily infected with this parasite may arrive too late at the spawning site to participate in mating. This could ensure a selection of 'good genes'.     

Vaccination of eels (Anguilla japonica and Anguilla anguilla) against Anguillicola crassus with irradiated L3

The original host of the swimbladder nematode Anguillicola crassus, the Japanese eel (Anguilla japonica) and the recently colonized European eel (Anguilla anguilla) were immunized with 40 irradiated (500 Gy) 3rd-stage larvae (L3) of this parasite and challenged with an infection of 40 normal L3. The immunization induced a significant reduction of the number of adult worms developing from the challenge infection in A. japonica, but not in A. anguilla. The induced resistance (calculated using the relation of the number of adult worms in immunized eels and in non-immunized control eels) in A. japonica was 87.3%+/-30.4%. Following a single infection, the percentage of adult worms found in A. japonica was lower as compared to A. anguilla, and the few adult worms were much smaller, revealing a lower susceptibility of A. japonica to A. crassus in comparison to A. anguilla. Both eel species developed an antibody response against A. crassus, but the level of antibody responses was not positively correlated with the protection against infection, suggesting that the antibody response is not a key element in resistance of eels against A. crassus. This study suggests that the original host of A. crassus is able to mount efficient protective immune responses against its parasite, whereas the newly acquired host seems to lack this ability.     

Distribution and prevalence of Anguillicola crassus in eels from the tidal Thames catchmentb

Data gathered between 1988 and 1992 document the spread of the parasitic nematode Anguillicola crassus among eels in the tidal Thames catchment. Eel samples revealed a parasite prevalence ranging between 12 and 32% with a variation in intensity of infection of between one and five nematodes per infected host. Differences in the salinity regime between sampling points may be linked to the range of levels of infection in eels because of the saline tolerance limits of parasite developmental stages. The euryhaline teleost, the smelt ( Osmerus eperlanus ) found throughout the tidal river has been shown by others to be able to transfer nematode larval stages experimentally to large eels. Smelt found in the tidal Thames thus could possibly act as a further intermediate host to the eel population. The results support the theories proposed by previous workers that the parasite originally entered the tidal Thames via the commercial trade in live eels.     

Spatial and temporal variation in Anguillicola crassus counts: results of a 4 year survey of eels in Mediterranean lagoons

We present the results of a survey of anguillicolosis in the Rh6ne River delta. From January 1997 to December 2000, a total of 13,319 eels (Anguilla anguilla from elver to silver phase) were examined, in which we found 22,227 swimbladder nematodes (Anguillicola crassus adults and pre-adults). A generalised linear model (GLM) framework was used to explore the relative contribution of various factors to the occurrence, intensity and abundance of the parasite. We reveal a major influence of the month of sampling, and we document the existence of a seasonal pattern with regular peaks in early summer and late winter. In contrast, the year of sampling is of secondary importance, and no particular trend in the development of the infection can be detected. More than a decade after the first record of A. crassus in the Rh?ne River delta, anguillicolosis has thus attained a constant infection rate of nearly 50%, with a mean number of 3 or 4 macroscopic lumen worms per infected eel. The eel length strongly influences the intensity and the abundance of the nematode, but has little if any effect on the probability of being infected. There exists a linear relationship between eel size and the number of parasites, but not between eel size and prevalence. We observe a decrease in the proportion of infected individuals among elver eels. We discuss this result in relation to the possible mortality of heavily infected individuals and/or a change in the eels' alimentary diet.     

Biased sex ratio in the European eel (Anguilla anguilla) swim-bladder parasite Anguillicola crassus, experimentally induced by 11-ketotestosterone

Parasites are intimately connected to the host in which they live, and some may be affected by the polluted environment of their host. The present study describes the effect of a steroid hormone (11-ketotestosterone) on the sex ratio of the invasive hematophagous nematode Anguillicola crassus Kuwahara, Niimi & Itagaki, 1974, when experimentally injected to European eels, Anguilla anguilla. Our results showed that this steroid induced a significant male-biased ratio in the nematode A. crassus infrapopulations, suggesting that the presence of endocrine disruptors in the environment may lead to skewed sex ratios among parasites.     

First record of Anguillicoloides crassus (Nematoda) in American eels (Anguilla rostrata) in Canadian estuaries, Cape Breton, Nova Scotia

In the summer of 2007, American eels, Anguilla rostrata, from 2 localities on Cape Breton Island, were found to be infected with the swim bladder nematode Anguillicoloides crassus. This is the first documented report of this highly invasive parasite in Canadian waters. More than half of the yellow eels in Mira River (6 of 10), and 1 eel (of 5) from Sydney Harbour were infected. Parasite intensity ranged from 1 to 11 worms per eel. The occurrence of A. crassus at these 2 localities suggests the need for a more extensive survey on the distribution of this exotic parasite in eel populations throughout Cape Breton Island.     

Changes in the levels of chloride cells and (Na+ + K+)-dependent ATPase in the gills of yellow and silver eels adapting to seawater.

Changes were measured in the numbers of chloride cells and the levels of (Na+ + K+)-DEPENDENT ATPase in the gills of immature, yellow eels and mature, silver eels during adaptation from freshwater to seawater. The percentage of chloride cells in yellow eels more than doubled after six days in seawater; at this time the specific activity and concentration of (Na+ + K+)-dependent ATPase in gills start to increase in parallel to reach maxima after two weeks that are 2.5 times the starting values. It is concluded that adaptation of yellow eels to seawater involves an increase in the numbers of chloride cells in gills as well as an increased amount of (Na+ + K+)-dependent ATPase per chloride cell. Mature silver eels in freshwater had essentially the same numbers of chloride cells and the same specific activity of the enzyme in the gills as yellow eels fully adapted to seawater. Transferring silver eels to seawater did not alter the percentage of chloride cells in gills although the level of (Na+ + K+)-dependent ATPase and its specific activity increased slightly. Thus, although the silver eel is better prepared for life in seawater than the yellow eel, it still has to attain an increased level of (Na+ + K+)-dependent ATPase in its chloride cells to be fully adapted to seawater.     

Switch in rod opsin gene expression in the European eel,Anguilla anguilla (L.).

The rod photoreceptors of the European eel, Anguilla anguilla (L.), alter their wavelength of maximum sensitivity (lambda max) from c.a. 523 nm to c.a. 482 nm at maturation, a switch involving the synthesis of a new visual pigment protein (opsin) that is inserted into the outer segments of existing rods. We artificially induced the switch in rod opsin production by the administration of hormones, and monitored the switch at the level of mRNA accumulation using radiolabelled oligonuleotides that hybridized differently to the two forms of eel rod opsin. The production of the deep-sea form of rod opsin was detected 6 h after the first hormone injection, and the switch in rod opsin expression was complete within four weeks, at which time only the mRNA for the deep-sea opsin was detectable in the retinal cells. It is suggested that this system could be used as a tractable model for studying the regulatory control of opsin gene expression.     

Structure and evolution of the teleost extraretinal rod-like opsin (errlo) and ocular rod opsin (rho) genes: is teleost rho a retrogene?

In Teleost fish examined to date the ocular rod opsin gene, rho, is intronless, unlike the rod opsin genes of other vertebrate classes which possess a five exon/four intron structure. We have examined in silico the structure of rho (which is expressed uniquely in the retina) and the closely related extraretinal rod-like opsin (exo-rhodopsin) gene, errlo (which is expressed uniquely in the pineal), in the puffer-fish, Fugu rubripes (Takifugu rubripes). Whilst the ocular rho is intronless in common with other Teleosts, the pineal errlo has the five exon/four intron structure common to the rod opsin gene of other vertebraes. A comparison of the sequence surrounding the errlo and rho loci indicates that the errlo locus is syntenic with RHO, the human rod opsin gene, rather than rho. We suggest that the intronless rho may have arisen through an ancient retrotransposition of a mature mRNA originating from errlo. This duplication event has occurred early in the evolution of the Actinopterygii (ray-finned fish) since the rho of the primitive Actinopterygians such as sturgeon, bowfin, and gar is also intronless. Since it appears that the intron containing errlo is the ancestral opsin gene that gave rise to the intronless rho in the Teleostei, errlo is therefore the true orthologue of the rod opsin gene in other vertebrate classes. We suggest that loss of expression of errlo in the retina could be related to the metabolic and physiological advantages, such as a reduction in splicing events during RNA processing, that may be conferred through possession of an additional, intronless rod opsin gene in the form of rho.     

The swimbladder nematode Anguillicola crassus in American eels (Anguilla rostrata) from middle and upper regions of Chesapeake Bay.

The patterns of infection of American eels Anguilla rostrata, with the introduced swimbladder nematode Anguillicola crassus, in tributaries of middle and upper Chesapeake Bay are described. A total of 423 subadult eels was collected from 8 Bay tributaries from spring 1998 to fall 1999. Also, 30 elvers were collected from Ocean City, Maryland, in spring 1998. The numbers of juvenile and adult specimens of A. crassus in the swimbladder wall and lumen were counted. No elvers were infected. In subadult eels, prevalence of adult and juvenile stages combined ranged from 13% to 82%; mean intensity ranged from 2.6 to 9.0 worms per eel. Infection levels were highest for Susquehanna River eels (northernmost river) and lowest in the southernmost sites: St. Jerome's Creek and the Pocomoke River. Although eels from these 2 localities were larger, the low infection rates there are most likely due to reduced transmission in higher salinity water and not to eel size. Eels with both adult and juvenile stages of A. crassus were more common than expected by chance. This might be explained by inhibition of juveniles migrating into the swimbladder lumen when adults are already present there.     

Humoral immune response of European eel Anguilla anguilla experimentally infected with Anguillicola crassus

A humoral immune response of the European eel Anguilla anguilla elicited by an experimental infection was demonstrated for the first time against the swimbladder nematode Anguillicola crassus. Eels were experimentally infected once or repeatedly and the antibody response was observed over a period of 325 d. Specific antibodies against A. crassus in the peripheral blood of the eels were measured using an ELISA and the immunoblot technique. Anti-A. crassus antibodies were first observed 8 wk post infection, and appeared to be independent of both the number of infective third stage larvae (L3) administered and the frequency of administration. However, individual eels showed great differences in the course of the antibody response. The late appearance of antibodies in the peripheral blood supports the hypothesis that not the invading L3 but rather the adult parasites elicit the production of specific antibodies. A stage-specific antibody response against the L3 was not observed. Main antigens are located in the body wall, especially in the gelatinous outer cuticle, of adult A. crassus.     

Dynamics and predicted decline of Anguillicola crassus infection in European eels, Anguilla anguilla, in Neusiedler See, Austria

The eel population in Neusiedler See has been maintained by regular massive stocking since 1958. After the establishment of the National Park Neusiedler See-Seewinkel in 1993, eel stocking was prohibited and the population, together with the specific parasites of eels, was predicted to decline to extinction within 10 years. This investigation was undertaken to document the decline and extinction of the Anguillicola crassus population in eels. From 1994 to 2001, 720 eels were collected from two sites in the lake. Prevalence and abundance of A. crassus were lower in spring than in summer and autumn and larger eels harboured more parasites than smaller ones. Neither year of study nor sampling site were correlated with parasite infection levels. No significant trend in the population parameters of A. crassus was detected over the 8 years of the survey. This suggested that there had been no significant decline in the eel population. This suggestion was confirmed by investigations of the fishery, which also found evidence of regular illegal stocking. The stability of the A. crassus population over the past decade seems to reflect the lack of change in eel population density. No mass mortalities of eels occurred over the period despite the many similarities between Neusiedler See and Lake Balaton in Hungary. Differences in eel size, eel diet and the lack of large-scale insecticide use are discussed as possible explanations for the absence of eel mass mortalities in Neusiedler See.     

Tissue-specific and developmental regulation of rod opsin chimeric genes in transgenic mice.

Chimeric gene fusions between 4.4 kb of rod opsin 5' flanking sequence fused to a diphtheria toxin gene and 4.4 kb or 500 bp of rod opsin 5' flanking sequence fused to the E. coli IacZ gene were used to generate transgenic mice for analysis of cell type-specific expression and temporal and spatial distribution of reporter gene product during retinal development. Opsin-diphtheria toxin transgene expression evoked photoreceptor-specific cell death. The 4.4 kb opsin-IacZ transgene followed temporal and spatial gradients of expression that approximate opsin expression. The 500 bp opsin fragment targeted expression to photoreceptors, but expression was weaker and nonuniform, suggesting that elements located upstream may be required for enhanced and uniform spatial expression.