Anguillicola crassus is the most invasive species of its genus and it is a successful colonizer of different eel species worldwide. It is so far the only species of the genus Anguillicola whose life cycle has been studied completely. To analyse whether differences in life cycle may explain differences in invasiveness, we infected European eels with Anguillicola novaezelandiae under laboratory conditions. Anguillicola novaezelandiae shows a synchronized development in the European eel. Eggs with second-stage larvae appeared 120 days after infection. No density-dependent effect in parasite development could be found for A. novaezelandiae. The present study shows that the life cycle of A. novaezelandiae differs on final host level compared with A. crassus in ways which result in a less successful invasion of new host species.

The effect of Anguillicola crassus (Nematoda) on gas composition in the swimbladder of eels (Anguilla anguilla) was studied using mass spectrometry in feral eels from two German rivers and in experimentally infected eels. In both naturally and experimentally infected eels significant correlations were observed between the proportion of oxygen in the swimbladder and level of infection with Anguillicola crassus. In swimbladders of naturally infected feral eels the contribution of oxygen to swimbladder gas was reduced by 36–62·9% and in experimentally infected eels it was reduced by 11·4–57% compared to uninfected controls. The proportion of CO2 appeared to be lower in infected swimbladders compared to uninfected ones. However, this change was not significant. The findings are discussed in relation to an altered structure of the swimbladder wall due to the parasite.

Hatching of eggs of Anguillicola crassus was studied at temperatures between 5 and 30°C. Hatching occurred between 10 and 30°C and the incubation time was inversely related to temperature. Survival, spontaneous and excitatory rate of activity of 2nd-stage larvae of A. crassus were also affected by temperature. The percentage of larvae eaten by Paracyclops fimbriatus was not always influenced by the activity of the larvae. Penetration efficiency into the haemocoel of the intermediate host was highest for freshly hatched larvae and declined gradually with age. Completely non-active larvae were still able to penetrate, although penetration efficiency was lower.

Density-dependent effects of Anguillicola crassus larval infections in the copepod intermediate host were examined experimentally. Three species of copepods (Cyclops vicinus, C. viridis and C. fuscus) were subjected to a range of doses of larval A. crassus within infection arenas. Prevalence, intensity and parasite dispersion (variance: mean abundance) values increase and then approach an asymptote as infection dose increases. Infection parameters differ between species of copepod. Increasing temperature has a negative effect on the establishment of the parasite population within the intermediate host. Parasite-induced host mortality increases with dose. These mechanisms have the potential to regulate populations of A. crassus larvae within the copepod population and hence the whole suprapopulation.

The parasitic nematode Anguillicola crassus was recently introduced into populations of the European eel, Anguilla anguilla. We investigated, under experimental conditions, the regulation of A. crassus infrapopulations. We tested the effects of (1) the resource-limited habitat of the parasite and (2) the coexistence of several developmental stages in its niche (the swim-bladder) on the composition of the infrapopulations. The results revealed that the respective effects of these factors differed substantially during the course of the infection. Third-stage larvae (L3s) establishment would not be constrained by the size of the swim-bladder. Their moult to fourth-stage larvae (L4s) would be accelerated as the number of L3s increased. The moulting time of L4s to adults would be reduced by males and would be constrained by the size of the swim-bladder. However, the moult of L4s to adults and their further development would be synchronized with those of the opposite sex. At the time of mating, the number of males and the body weight of adults would depend on the size of the swim-bladder. Soon after the laying of eggs, the developmental constraint on the late L3s would decrease. When adults die, constraints would cease and late larval stages would moult to become adults.

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.

To investigate whether the stress response of European eels infected with Anguillicola crassus is influenced by environmental pollutants, experimentally infected eels were exposed to Cd and/or to 3,3′, 4,4′, 5-pentachlorobiphenyl (PCB 126). Serum cortisol and glucose concentrations of these eels were monitored over a period of 103 days and were compared with data from infected, unexposed eels as well as with data from uninfected eels. Additionally, the levels of cortisol were correlated with concentrations of Anguillicola-specific antibodies. All eels showed an initial increase of the cortisol levels until day 63. This general elevation of plasma cortisol is most likely due to handling stress, as all eels were repeatedly netted and afterwards inoculated with a feeding tube. At the end of the exposure period eels which were infected and those which were infected and simultaneously exposed to Cd and PCB showed significantly higher levels than the controls. The general course of serum glucose levels in eels resembled that of cortisol. Accordingly, Spearman correlation analysis revealed that an increase in serum cortisol concentrations is correlated with rising levels of glucose. With respect to immune-endocrine interactions a significant negative correlation between cortisol and anti-A. crassus antibodies was found. Our data show that A. crassus is the most potent stressor for European eels among the treatments tested within this study. This is important in terms of ecotoxicological studies as the main effects are caused by parasites rather than chemicals. Accordingly, effects of parasites on the physiological homeostasis of organisms must be considered in ecotoxicology. From the parasitological point of view our results suggest that probably as part of an unbalanced host-parasite interaction A. crassus evokes a strong cortisol response in A. anguilla, thereby suppressing the immune response which in turn enables the parasite to establish. The parasite-induced stress response in the newly adopted European eel might be one of the factors which contributes to the extremely effective colonizing strategy of A. crassus.

European eels Anguilla anguilla from the rivers Thames and Test, in the south of England, were examined between 2000 and 2003 for infection with the swim-bladder nematode Anguillicola crassus. Since its introduction to Thames eels at tidal estuarine locations circa 1987, A. crassus has become established in non-tidal freshwater stretches upriver and data from these locations are reported for the first time. The prevalence of infection at Thames estuary locations was higher during 2000–2003 than for the period 1987–1992. By 2003, similar prevalences were observed at freshwater and estuarine locations, but infection intensities were significantly higher in freshwater. Eels from the river Test appear to have been recently colonized by A. crassus (circa 2000). Parasite population establishment within these eels was uncharacteristically slow during 2000–2001, with low prevalence and intensity of infection, and few gravid females during this period. By 2003, infection levels in Test eels were similar to those in Thames eels. The expansion of the A. crassus population in Test eels has occurred in the absence of the paratenic host ruffe Gymnocephalus cernuus, and at suboptimal pH for the survival and infectivity of free-living larvae. The epidemiology of A. crassus in Test eels demonstrates that transmission of A. crassus by ruffe is not required for high prevalence and intensity of infection in eels. However, the consistently low and atypical levels of infection in Test eels during 2000 and 2001 suggests that paratenic transmission by ruffe may provide a substantial contribution to the dynamics of A. crassus in eels in the early years following introduction, by facilitating the rapid increase in prevalence and intensity of A. crassus infection that typically occurs during this time.

The swim-bladder nematode Anguillicola crassus originates from the Far East where it is a parasite of the Japanese eel (Anguilla japonica). After A. crassus was introduced to Europe, it became a predominant parasite of the European eel (Anguilla anguilla). A study performed with experimentally infected eels (98 days, 23 °C) revealed significant differences in the susceptibility of the two eel species to this parasite. The recovery rate of 30 administered infective A. crassus larvae (L3) from A. japonica was less than half of that from A. anguilla (33·2% and 13·8%, respectively). Almost 60% of the worms recovered from A. japonica were found as dead, encapsulated and necrotic larvae in the swimbladder wall. In contrast, no dead larvae were found in A. anguilla. Additionally, the development of the worms was shown to be significantly slower in A. japonica compared with A. anguilla. The lower survival rate of the worms, together with their slower development, resulted in a significantly lower adult worm burden (11 and 428 mg wet weight, respectively) and in a decreased reproductive success in A. japonica compared with A. anguilla. These results demonstrate that the original host, A. japonica, possesses more effective defence mechanisms against A. crassus than does the non-adapted host, A. anguilla.

We have developed an index based on macroscopic criteria to easily assess the health state of the eel swimbladder as a result of infection by the nematode Anguillicola crassus. In the sampling area (brackish lagoons of the French Mediterranean coast), 92% of the host sample (1251 eels covering all size classes) showed pathological signs of infection. The general trend was for increasing damage as eel size increased, thus suggesting accumulation of pathological effects. We also revealed a non-linear relationship between the swimbladder index and the abundance of living worms. In particular, we showed that severely damaged swimbladders harboured very few living nematodes. We argue that the swimbladder degenerative index more closely reflects the parasite pressure than does classic parasite count. We found seasonal variation in the swimbladder index, with maximum damage occurring in July and thereafter a trend for healthy individuals. We discuss the possibility that the seasonal decrease in the swimbladder index could reflect the death of the more severely affected individuals during the warmest months.

The study was designed to test the hypothesis that the majority of individuals in a population of Anguillicola crassus can tolerate seawater by osmoconformation with the blood plasma of the eel host. The osmolality of the pseudocoelomic fluid of pre-adult and adult A. crassus was compared to the osmolality of eel plasma after eels were maintained in laboratory freshwater for 2 weeks (short-term transfer) or 3 months (long-term transfer) or in natural seawater for 2 h (acute transfer), 2 weeks or 3 months. The majority of A. crassus (at least 90% of the tested population) osmoconform with their hosts in seawater within ±30 mOsm/kg of host osmolality. Some pre-adults and adults (15–21% of the total population) were unable to withstand osmotic stress resulting in vacuolation of the hypodermis and intestinal wall, and cuticular detachment. The reasons for variation in the tissue tolerance of A. crassus to increased osmolality of host plasma are unknown and are not related to maturity, sex or conditions in the swimbladder. Osmoconformation in the majority of the population, however, enables parasites to survive and reproduce for long periods in seawater eels. The adaptation of A. crassus to its euryhaline host has played an important part in the rapid spread of the swimbladder nematode in populations of the European eel.

The purpose of this study was to determine under laboratory conditions over a period of 311 days if infection with the nematode Anguillicola crassus induces stress in European eels (Anguilla anguilla), and stimulates the endocrine stress axis as measured by serum cortisol levels. Eels were experimentally infected with 3rd-stage larvae (L3) in different doses to simulate natural conditions with varying infection pressures. Blood samples were drawn from the caudal vein every 2 weeks and serum cortisol concentrations were determined by radioimmunoassay (RIA). The results showed that the application of L3 resulted in a significant increase in the cortisol levels. The period of time at which elevated cortisol values were observed was consistent with the time of larval development and the appearance of adult A. crassus. Thus, there is a stress response to the larval and young adult stages, but no chronic response to older adults. Therefore, it is likely that infection of eels under natural conditions especially with a high number of larvae may be a considerable stressor, in combination with different environmental factors like water temperature, pH, oxygen concentration, pollution and inter-individual relationships.

The ability of the nematode Anguillicola crassus to infect eel larvae (glass-eel stage) was tested. The results show that glass-eels fed on infected copepods, the natural intermediate host of the nematode, can be infected. Light microscopical examination of the infected developing swimbladder tissue revealed that the infection results in a significant thickening of the connective tissue. The basolateral labyrinth of gas gland cells is very much reduced in infected swimbladders, and the distance of gas gland cells to blood capillaries is enlarged. Critical swimming speed, defined as the speed where the larvae were no longer able to swim against the current, was similar in infected and uninfected animals. At intermediate speeds (about 60–80% of critical swimming speed) infected eels showed a slightly higher swimming activity than control animals. Resting oxygen consumption, measured as an index of metabolic activity, within the first 2 months of infection was higher in control animals, which may be due to a reduced rate of activity in infected glass-eels. By 4–5 months after the infection, however, it was significantly higher in infected animals. This may indicate that at this stage a higher activity of the animals is required to compensate for the increase in body density, but swimming performance of infected and non-infected glass-eels was not significantly different. Oxygen consumption during swimming activity, measured in a swim tunnel at 50% of maximal swimming speed, also was not affected. The results thus show that even glass-eels can be infected with A. crassus, and this probably contributes to the rapid spread of the nematode in Europe. While aerobic metabolism during swimming activity is not affected at this stage of infection, the swimbladder tissue shows severe histological changes, which most likely will impair swimbladder function.

The study aims were to quantify the survivorship of Anguillicola crassus in eels in seawater and investigate transmission in estuarine (50% seawater) and marine (100% seawater) simulated conditions. Most A. crassus were able to survive and reproduce in 50% and 100% seawater eels for up to 6 months and therefore could survive during the spawning migration of eels to the Sargasso Sea. Up to 10% of the parasite metapopulation were damaged after long-term exposure to 50% or 100% seawater. Transmission of A. crassus was completed in 50% and 100% seawater in the laboratory, although infection levels in 100% seawater hosts were always lower. Transmission in estuaries is ecologically possible since the copepod Eurytemora affinis is susceptible to infection and is the dominant autochthonous species in most tidal estuaries in the Northern hemisphere. Transmission at sea is unlikely due to lack of suitable intermediate hosts. The data support the suggestion that dissemination of A. crassus has been facilitated by natural movement of eels in coastal waters. It is possible that A. crassus could impair the success of the eel spawning migration to the Sargasso Sea, although there is no direct evidence to support this.

The effect of the parasitic swimbladder nematode, Anguillicola crassus, on the physiological status of wild European eels (Anguilla anguilla) was investigated during an 18 month survey (February 1995 until September 1996), and compared with that of wild uninfected eels collected over the same time-period. Despite the occurrence of up to 15 blood-feeding adults in the swimbladder lumen of the infected eels and as many as 25 additional larvae in the swimbladder wall, there were no major differences in hormonal, metabolic or osmoregulatory status of the 2 groups of eels. Wild European eels appear to adapt to chronic parasitism with Anguillicola crassus. The possible adverse effects of additional simultaneous stressors are discussed.

A total of 121 European eels (Anguilla anguilla) from 2 sampling sites on the River Rhine were investigated in respect of their parasite communities. Special attention was given to the swim bladders, intestines, gills and fins of the fish. Twelve different parasite species were found to live in and on the eels. Data from each sampling site were kept separate. Parasites found in descending order of prevalence were: Anguillicola crassus, Trypanosoma granulosum, Myxobolus sp., Paratenuisentis ambiguus, Pseudodactylogyrus sp., Bothriocephalus claviceps, Myxidium giardi, Pomphorhynchus laevis, Trichodina sp., Raphidascaris acus, Acanthocephalus lucii and Acanthocephalus anguillae. Significantly different prevalences were reported for L3 larvae of A. crassus, adult P. ambiguus, B. claviceps and Myxobolus sp. at the 2 sampling sites. The highest number of parasite species was recorded from the intestine, which contained up to 6 different helminths. The coexistence of the acanthocephalans P. laevis and P. ambiguus, which showed clear patterns of distribution within the intestine of the respective hosts, was reported for the first time. Up to 3 different helminth species were found in the intestine of individual fish. Among those, acanthocephalans were the most prevalent worms with the eel-specific parasite P. ambiguus as the dominant species not only of the intestinal but also of the total component communities. Both infra and component communities exhibited low diversity and were dominated by this single species. The evenness reached only approximately 50% or less and it remained unclear why the helminth communities of the eels from the River Rhine with its huge catchment area exhibit such a low parasite diversity and high dominance.

Density-dependent effects on adult and larval stages of the introduced nematode parasite Anguillicola crassus in the European eel definitive host were examined in the laboratory in naturally infected fish, and in field samples. The effect of adult nematode subpopulations on larval development over time and the effect of increasing adult intensity on the gravid female subpopulations were investigated. At high adult subpopulations the movement of larvae from the swimbladder wall into the swimbladder lumen appears to be inhibited, and A. crassus larvae to be arrested in development in a density-dependent manner. The number of gravid females per host reaches a constant level, and so the proportion of gravid female nematodes per adult subpopulation decreases relative to further increasing total adult nematode numbers. Both these mechanisms have the potential to regulate infrapopulations of A. crassus within the eel definitive host and thus the parasite suprapopulation.

The larval stages of Anguillicola crassus, a parasite of the swimbladder of eels, are described, using light and scanning electron microscopy. Second-stage larvae in copepods are characterized by a globular cephalic end and third- and fourth-stage larvae by an apical selerotized ornamentation consisting of two U-shaped median pieces. Third- and fourth-stage larvae are difficult to distinguish from each other using light microscopy because the third moult has never been observed and there are no obvious characters differentiating the two stages. Three characters, however, observed by using high magnification of light and scanning electron microscopy are considered as distinctive between the two stages, namely presence or absence of deirids and lateral alae and differences in the structure of the cuticle. These morphological characters will permit epidemiological studies at the larval level.