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'.     

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.     

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.     

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.     

Occurrence of Anguillicola crassus (Nematoda: Dracunculoidea) in Japanese eels Anguilla japonica from a river and an aquaculture unit in SW Taiwan

The infection by swimbladder nematodes of the genus Anguillicola (Dracunculoidea: Anguillicolidae) was examined in 2 populations of the Japanese eel Anguilla japonica in SW Taiwan. Wild eels from the Kao-Ping river were compared with cultured eels from an adjacent aquaculture unit. Only the cosmopolitan species Anguillicola crassus was present. Among wild eels, prevalence of infection varied between 21 and 62%, and mean intensity between 1.7 and 2.7 for adult worms. Similar intensity values (1.3 to 2.8) were recorded for the larvae. In cultured eels, prevalence as well as mean intensities were higher. In the cultured hosts, mean larval intensities exceeded those of adult worms 2-fold, and maximum larval intensities were 4- to 5-fold higher than in eels from the river. In cultured eels, dead larvae were also more abundant than in wild eels. We conclude that infrapopulations of A. crassus in Japanese eels are regulated by the defense system of this host, intraspecific density-dependent regulation being less likely as the major regulatory mechanism. No influence of the parasite on eel condition was found in either wild or cultured eels, indicating a low or moderate pathogenic effect of A. crassus on this host. This study shows that A. crassus is moderately common in cultured and wild Japanese eels in Taiwan, where the parasite is endemic.     

Spatio-temporal dynamics of the parasitic nematode Anguillicola crassus in Flanders, Belgium

Despite Egusa's earlier warning of the damage that the parasitic nematode Anguillicola crassus could inflict on the European eel Anguilla anguilla, its introduction in Europe was a fact in the early 1980s. Based on an elaborate dataset on Anguillicola crassus infection of 11 river catchments, this paper presents the results of a detailed study on the dispersal of the parasite in Flanders, Belgium, and the host-parasite relationship. In addition, data from 1986 and 1997 are used for comparative purposes, providing a perspective on the temporal infection pattern over 15 yr. The presence of A. crassus in Flanders was first discovered in 1985; 2 yr later a survey revealed a prevalence of 34.1% and a mean infection intensity of 5.5, based on adult nematodes only, and 10 yr later the parasite was present at all 11 sites sampled. Prevalence had increased to 62.5 % but the mean infection intensity had decreased to 3.9 adults per infected eel. Finally, in the year 2000, a third study revealed that A. crassus was present in 139 of 140 investigated sites; a further increase in prevalence to 68.7% and a decrease in mean infection intensity to 3.4 adults per infected eel was observed. When all larval stages were taken into account, mean prevalence amounted to 88.1% and mean intensity to 5.5 adults. The high infection level in Flanders is thought to be the result of restocking with glass eel and yellow eel, both of which are susceptible to A. crassus. The general infection parameters were similar in all 11 river catchments. It is possible that in Flanders both prevalence and mean infection intensity are stabilizing due to density-dependent regulation of the parasite infrapopulation. Fibrotic swimbladder walls were observed, mainly in large eels, and 20% of the total number of nematodes consisted of encapsulated larvae in the surveys of 1997 and 2000; 8 cases of swimbladder regeneration were observed.     

Impacts of the swimbladder nematode Anguillicola crassus on Anguilla anguilla: variations in liver and spleen masses

Variations in the liver and spleen masses of the eel Anguilla anguilla were analysed in relation to the parasite load of Anguillicola crassus at autopsy (current infection by swimbladder lumen worms) and in relation to the severity of damage observed in the swimbladder (a way of assessing the intensity of past infections). None of these measures of parasite pressure were shown to account for variation in the relative liver mass, either when controlling for somatic mass or eel age. In marked contrast, a significant increase in spleen size was revealed in eels harbouring many lumen worms and also in eels with severe damage in the swimbladder. Splenic enlargement was nearly two‐fold higher among severely affected eels (harbouring more than seven lumen parasites and showing severe damage in the swimbladder) than among infection‐free eels (no lumen parasites and no pathological signs in the swimbladder). Several possible hypotheses are reviewed before arguing for an adaptive host response involving the haematological and immunological functions of the spleen. Indeed, among eels with no pathological signs in the swimbladder, the relative spleen mass was positively associated with the mass of lumen parasites, which suggests a hyper‐synthesis of blood cells by the spleen in response to the bloodsucking activity of lumen worms. Nevertheless, among eels with no lumen parasites at autopsy, there was still an increase in spleen size in relation to the severity of the swimbladder damage, which also suggests a hyper‐synthesis of splenic immune cells (lymphocytes and macrophages) in reaction to damaged tissues and particularly to larvae in the swimbladder wall.     

Development of Anguillicola crassus (Dracunculoidea, Anguillicolidae) in experimentally infected Balearic congers Ariosoma balearicum (Anguilloidea, Congridae).

The development of Anguillicola crassus in experimentally infected Ariosoma balearicum (Anguilloidea, Congridae) kept in seawater was studied in the laboratory. In parallel trials the effect of water salinity on the development of larval A. crassus in European eels Anguilla anguilla was also investigated using eels kept in seawater of a salinity of 34 per thousand. Both eel species were orally inoculated with L3 larvae of A. crassus and then maintained for up to 3 mo at 18 degrees C in seawater. 110 d post infection, no adult but larval (L3 and L4) stages of A. crassus were detected in the swimbladder wall of Balearic congers, although this period of time was sufficient for the parasites to develop to the adult stage in European eel kept in seawater. The results presented suggest that the definitive host specificity of A. crassus comprises species of the family Anguillidae (i.e. the genus Anguilla), but not members of the Congridae. Theoretically however, A. balearicum might serve as a metaparatenic host. Factors determining the definitive host range of A. crassus remain to be elucidated. Water salinity does not seem to act as a factor affecting definitive host specificity once the parasite has become ingested by the eel.     

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.     

Anguillicola crassus (Nematoda, Dracunculoidea); first documented record of this swimbladder parasite of eels in Ireland

In summer 1998 European eels infected with the swimbladder nematode Anguillicola crassus were detected in the Erne catchment, Ireland, probably introduced recently through the commercial eel trade.     

Histopathological changes in the swimbladder wall of the European eel Anguilla anguilla due to infections with Anguillicola crassus

The histopathological changes in swimbladders of European eels naturally and experimentally infected with Anguillicola crassus were studied using transmission and scanning electron microscopy. During the course of probably several infections swimbladders undergo characteristic changes. In addition to the thickening of the entire swimbladder wall, and to the folded internal surface of this organ, inflammation, migration of white blood cells, fibrosis and changes in the epithelial cells are frequently seen. Epithelial cells tend to proliferate heavily and form hyperplastic tissues; these processes are accompanied by changes in the internal structure of the cells. The normally cubic cells become spherical or columnar and form folds facing the lumen of the swimbladder. As a consequence, most of these cells lose contact with the blood vessels and show no strict polarity. In heavily affected swimbladders the basal labyrinth of the epithelial cells is reduced, i.e. becomes shorter and less densely packed. The lamina propria shows severe fibrosis with infiltration of white blood cells. Larvae of A. crassus, inhabiting the wall of the swimbladder, were found to be surrounded by cell debris, but this local necrosis does not affect the entire swimbladder in its overall structure. These histological findings can partly explain changes in the gas composition in eels infected with A. crassus.     

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.     

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.     

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.     

Comparative study of X-ray computerised tomography and conventional X-ray methods in diagnosis of swimbladder infection in eels caused by Anguillicola crassus

To date, swimbladder lesions due to Anguillicola crassus infection of the European eel Anguilla anguilla have so far been studied only by conventional X-ray methods. This is the first study to report the use of computerised tomography (CT) for studying lesions induced by anguillicolosis. Of 50 eels caught by electrofishery from Lake Balaton, Hungary, in autumn 2002 and pre-selected by a conventional X-ray method, 22 specimens were examined with a Siemens Somatom Plus S40 spiral CT scanner. Tomograms, radiographs and photographs of 5 of these, showing anguillicolosis-induced swimbladder lesions of varying severity, are presented. Computerised tomograms provide information on the inner structure, air content and wall thickness of the swimbladder as well as on the number of worms it contains. When the swimbladder is not severely affected or not completely filled with worms, computerised tomography provides adequate data on the shape of the swimbladder, thickness of the swimbladder wall and the location of worms in the lumen. However, in more severe cases, i.e. when the swimbladder is tightly packed with worms or contains no air as a result of wall-thickening, this method fails to determine the number and location of helminths or the thickness of the swimbladder wall.     

An invasion record for the swimbladder nematode Anguillicoloides crassus in European eel Anguilla anguilla in a deep warm-monomictic [corrected] lake, from invasion to steady state

This study is the first account of the establishment and development of the neozoic nematode parasite Anguillicoloides crassus in its host, the European eel Anguilla anguilla, in a deep, warm-monomictic [corrected] lake. A 21 year study of A. crassus took place in Upper Lake Constance (ULC), Europe's second largest pre-alpine lake. The study included two extensive surveys, one in 1991 during the initial parasite invasion phase and the second in 2006 when the infection was well established. The subtropical swimbladder nematode A. crassus was first recorded in A. anguilla in ULC in 1989. Prevalence reached 60% in 1992 and remained at this level until 2007. In 2008, prevalence decreased to 48%. Infection intensity peaked in 1993 at a mean value of 16 adult parasites per host fish. Around 90% of all A. anguilla examined displayed swimbladder lesions, with a significant trend to increasing severity over time. Moreover, heavy swimbladder lesions were seen in c. 10% of A. anguilla ready to migrate to their spawning habitat. Both ruffe Gymnocephalus cernuus and sunfish Lepomis gibbosus serve as paratenic hosts for A. crassus in ULC. Gymnocephalus cernuus seems to be the main vector, and infection is especially frequent in spring possibly caused by reduced immune system efficacy of G. cernuus during winter. In 1991, hypochromic anaemia was prevalent in ULC A. anguilla acutely infected with A. crassus, whereas in 2006 blood values were indicative of chronic infection. The growth and survival rates of A. anguilla during their continental phase were not noticeably altered in infected fish, but damage to the swimbladder probably impairs migration potential and thus the subsequent breeding success of the oceanic phase.     

Physiological responses to acute temperature increase in European eels Anguilla anguilla infected with Anguillicola crassus

The swimbladder parasite, Anguillicola crassus has infected, and spread rapidly, through European eel Anguilla anguilla (L.) populations over the past 20 to 25 yr. Our aim in the present studies was to elucidate whether the presence of A. crassus in these eels alters their rapid physiological responses to an acute temperature increase, compared to the response of uninfected fish. Both infected and uninfected fish showed significant increases in plasma cortisol after 2 h at a raised environmental temperature with increased plasma glucose after 6 h. However, infected eels exhibited a slight lag in glucose mobilisation, which may be due to the metabolic cost of harbouring a sanguiverous parasite. Both infected and uninfected fish showed a significant increase in haematocrit after 6 h of temperature elevation, but only uninfected fish exhibited a significant increase in haemoglobin at this point. However, there were no significant changes in mean erythrocyte haemoglobin concentration in either group. Our results suggest that acute temperature increase alone is unlikely to cause significant mortality of A. crassus-infected European eels; however, the effects of chronic increases in temperature in combination with other factors such as toxicants and hypoxia requires examination.     

The spread of the eel swimbladder nematode Anguillicola crassus through the Erne system, Ireland

The spread of Anguillicola crassus was documented and showed an increase in both prevalence (9·9%) and mean intensity (6·7). Infected eels Anguilla anguilla were recorded 30 km downstream of the first recorded sites of infection. Migrating silver eels from commercial nets near the outlet of the Erne during October-December 1999 were also infected. Continued spread of A. crassus through Ireland's major wild eel fisheries now appears likely.     

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.     

Anguillicolosis: dynamics of the infection over two decades

The dynamics of the infection of the European eel Anguilla anguilla L. by the Asian nematode Anguillicola crassus Kuwahara, Niimi and Itagaki, 1974 (i.e. anguillicolosis) was monitored over 2 decades in an oligohaline canal in southern France (Camargue, Mediterranean coast). Since the first mention of the parasite in this canal in 1985, which was also the first record in France, prevalence of pre-adult and adult forms has risen from 32 to 73%. However, during the last 7 yr (1997 to 2003), prevalence seems to have stabilized around values of 60 to 70% and parasite load, though inter-annual variation is substantial, shows no sign of increase (intensity for the last 5 yr: min. = 3.70, max. = 9.66, mean = 6.01). Our results thus confirm the dynamic pattern observed elsewhere in Europe, i.e. a rapid spread following the introduction of the parasite in a water system and then stabilization around ceiling levels. We review possible mechanisms that may explain such a leveling off in the infection spread. We particularly document the possibility that repetitive infections may render the infected organ, i.e. the swimbladder, unsuitable for further A. crassus establishment. In support of this hypothesis, we showed that the infection rate is lower among eels with severely damaged swimbladders.