Encapsulation of Anguillicola crassus reduces the abundance of adult parasite stages in the European eel (Anguilla anguilla).

Encapsulation of the parasitic nematode Anguillicola crassus Kuwahara, Niimi & Hagaki is commonly observed in its native host, the Japanese eel (Anguilla japonica Temminck & Schlegel). Encapsulation has also been described in a novel host, the European eel (A. anguilla L.), and there is evidence that encapsulation frequency has increased since the introduction of A. crassus. We examined whether encapsulation of A. crassus provides an advantage to its novel host in Lake Müggelsee, NE Germany. We provide the first evidence that encapsulation was associated with reduced abundance of adult A. crassus. This pattern was consistent in samples taken 3 months apart. There was no influence of infection on the expression of the two metabolic genes studied, but the number of capsules was negatively correlated with the expression of two mhc II genes of the adaptive immune response, suggesting a reduced activation. Interestingly, eels that encapsulated A. crassus had higher abundances of two native parasites compared with non-encapsulating eels. We propose that the response of A. anguilla to infection by A. crassus may interfere with its reaction to other co-occurring parasites.

Molecular evidence linking the larval and adult stages of Mexiconema cichlasomae (Dracunculoidea: Daniconematidae) from Mexico, with notes on its phylogenetic position among Dracunculoidea.

We describe the larval developmental stages and life cycle of the dracunculid nematode Mexiconema cichlasomae in both the intermediate, Argulus yucatanus (Crustacea: Branchiura), and definitive hosts, Cichlasoma urophthalmus (Perciformes: Cichlidae), from the Celestun tropical coastal lagoon, Yucatan, Mexico. The morphological analyses showed significant differences between the total length of L1 found in M. cichlasomae gravid female and L2-L3 in A. yucatanus. This result indicates that the M. cichlasomae larval development occurs in the intermediate host. We obtained sequences from the small subunit (SSU) ribosomal marker from larval stages of M. cichlasomae in A. yucatanus and adult nematodes in C. urophthalmus. Our morphological and molecular results support conspecificity between M. cichlasomae larvae in A. yucatanus and the adult stages in C. urophthalmus. We briefly discuss the phylogenetic position of M. cichlasomae among the Daniconematidae, and provide evidence of the monophyly of the daniconematids associated with branchiurid intermediate hosts. Based on the phylogenetic results, we support the transfer of the Mexiconema genus to the family Skrjabillanidae and do not support the lowering of family Daniconematidae to subfamily.

Effects of Anguillicola novaezelandiae on the levels of cortisol and hsp70 in the European eel.

The nematodes Anguillicola novaezelandiae and Anguillicola crassus are both alien parasites of the European eel with severe adverse effects on their new host. Both species differ in terms of their invasiveness and their severity of harmful effects on the European eel. The purpose of this study was to determine under laboratory conditions whether stages of A. novaezelandiae induce stress in European eels (Anguilla anguilla) and if these levels differ from stress levels induced by A. crassus. We analysed levels of plasma cortisol and hepatic hsp70 of eels experimentally infected with A. novaezelandiae and compared them to uninfected eels as well as to eels experimentally infected with A. crassus. Larval stages of A. novaezelandiae induced higher levels of plasma cortisol compared to uninfected controls, while adult parasites increased the levels of hepatic hsp70 above those of uninfected controls. The eels' cortisol response is induced by larval stages of A. novaezelandiae, while adult stages elevate levels of hepatic hsp70. Levels of stress induced by A. novaezelandiae are comparable to those induced by A. crassus.

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.

Philometrids (Nematoda: Philometridae) in carangid and serranid fishes off New Caledonia, including three new species.

A recent examination of newly obtained specimens of philometrid nematodes (Philometridae) parasitising carangid and serranid fishes off New Caledonia, South Pacific, revealed the presence of several nematodes of the genus Philometra Costa, 1845, including three new species: P. austropacifica n. sp. (males and females) from the ovary of Alepes vari (Carangidae), P. piscaria n. sp. (males) from the ovary of Epinephelus coioides (Serranidae), and P. selaris n. sp. (males) probably from the abdominal cavity (found in washings) of Selar crumenophthalmus (Carangidae). The new species are characterised mainly by the length and structure of the spicules and gubernaculum, body size, their location in the host and the type of host. Philometra austropacifica n. sp. is the first known nominal gonad-infecting species of Philometra parasitising a carangid fish. In addition, the gravid female of P. fasciati Moravec & Justine, 2008 from the ovary of Epinephelus fasciatus (Serranidae) is described for the first time. Carangid host fish were identified by both morphology and DNA barcoding.

Temporal variation of Mexiconema cichlasomae (Nematoda: Daniconematidae) in the Mayan cichlid fish Cichlasoma urophthalmus and its intermediate host Argulus yucatanus from a tropical coastal lagoon.

The aim of the present investigation was to determine whether temporal variation in environmental factors such as rainfall or temperature influence long-term fluctuations in the prevalence and mean abundance of the nematode Mexiconema cichlasomae in the cichlid fish Cichlasoma uropthalmus and its crustacean intermediate host, Argulus yucatanus. The study was undertaken in a tropical coastal lagoon in the Yucatan Peninsula (south-eastern Mexico) over an 8-year period. Variations in temperature, rainfall and monthly infection levels for both hosts were analysed using time series and cross-correlations to detect possible recurrent patterns. Infections of M. cichlasomae in A. yucatanus showed annual peaks, while in C. urophthalmus peaks were bi-annual. The latter appear to be related to the accumulation of several generations of this nematode in C. urophthalmus. Rainfall and temperature appear to be key environmental factors in influencing temporal variation in the infection of M. cichlasomae over periods longer than a year together with the accumulation of larval stages throughout time.

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.

Prevalence, intensity, and differential development of Pseudodelphis oligocotti (Nematoda: Dracunculoidea) in sympatric fish hosts of the northeastern Pacific coast.

Counter to expectations of coevolved parasite-host relationships, parasites frequently infect hosts that never contribute to their reproduction, making the identification of a parasite's true host-specificity problematic. Pseudodelphis oligocotti (Nematoda: Dracunculoidea) infects several coastal Pacific fishes, but its course of development appears highly variable, suggesting that incidence does not reflect effective host range. To determine the host range of P. oligocotti and describe its relationship to various potential hosts, 24 fish species were examined from several British Columbia localities for prevalence, intensity, and extent and tissue location of parasite development. Pseudodelphis oligocotti infects 9 species of fishes from 5 orders, of which penpoint gunnel, Apodichthys flavidus, showed the highest prevalence and intensity, up to 80% and 19 (+/- 17.1 SD) worms per host, respectively. Although subadult and adult P. oligocotti occurred in all 9 fishes, larvigerous P. oligocotti only occurred in A. flavidus and rarely in the northern clingfish, Gobiesox maeandricus. Infective first-stage larvae were recovered from gill tissue of A. flavidus. Thus, at most only 2 of the 9 host species infected by P. oligocotti actually contribute to its transmission. The occurrence of P. oligocotti in diverse hosts may be accounted for by the parasite's indiscriminant mode of transmission via ingestion of free-living intermediate copepod hosts, where highly exposed or more suitable fishes (or both) are closely related by diet and microhabitat. This study demonstrates how parasite transmission and host ecology can greatly affect observed host range and ultimately its potential for expansion.

Some aspects of the taxonomy and biology of dracunculoid nematodes parasitic in fishes: a review.

The nematode superfamily Dracunculoidea includes 166 recognized species, of which 150 (90%) are parasitic in about 300 species of freshwater, brackish-water and marine fishes. Fish dracunculoids are placed in 31 genera (86% of all dracunculoid genera) belonging to eight of the nine dracunculoid families: Anguillicolidae, Daniconematidae, Guyanemidae, Lucionematidae, Micropleuridae, Philometridae, Skrjabillanidae, and Tetanonematidae; the genus Lockenloia is considered incertae sedis. Because of difficulties in studying fish dracunculoids, associated with their morphological and biological peculiarities, most species of these largely histozoic parasites are poorly known and males of the majority of species and of eight genera have not yet been discovered. It is apparent that the present classification system of dracunculoids as a whole does not reflect phylogenetic relationships and a taxonomic revision of this nematode group, based on detailed morphological (including SEM and TEM), life history and molecular studies of individual species, is quite necessary. Data on the biology of fish dracunculoids is scarce. In known cases, their life cycles involve copepods, ostracods or branchiurids as intermediate hosts and, sometimes, fish paratenic hosts are known to occur in dracunculoid species parasitizing as adults piscivorous definitive hosts. However, nothing is known about the life cycles of representatives of 20 genera. Some species of dracunculoids, particularly of philometrids, are highly pathogenic and are known as agents of serious fish diseases. During recent years, especially the importance of Philometra spp. parasitizing the gonads of many species of marine fishes has increased due in particular to the rapid development of marine aquaculture, because they may significantly decrease fish reproduction or even cause full parasitic castration. Therefore, further detailed studies on fish dracunculoids are significant not only from the theoretical viewpoint, but they may also have practical implications.

[Pseudorascora parva (Teleostei, Cyprinidae), an invasive species, a new vector for the maintenance and dissemination of anguillicolosis in France?]]. / Pseudorasbora parva (Téléostéen, Cyprinidae) espèce invasive, nouvel agent du maintien et de la dissémination de l'anguillicolose en France?

Pseudorasbora parva (Pisces, Cyprinidae) is an invasive species from eastern Asia. Known in France since 1983, it has only been observed since 1993 in the Rhône delta where large populations occur. 203 specimens of that fish species have been examined for L3-larvae of Anguillicola crassus. 35% of the fish were found to be infected.

Effect of salinity on hatching, survival and infectivity of Anguillicola crassus (Nematoda: Dracunculoidea) larvae.

The effect of salinity on hatching, larval survival and infectivity of Anguillicola crassus was studied under experimental conditions using eggs obtained from naturally infected eels. Egg hatching rate, second-stage larval survival and larval infectivity were maximal in fresh water and declined with increase in salinity. Larvae survived up to 100 d in fresh water, 70 d in 50 % sea water and 40 d in 100% sea water. Infectivity experiments demonstrated that salinity influenced transmission success throughout the life cycle by decreasing total infectivity of the larval population in utero within female A. crassus and when larvae were free-living in the aquatic environment. Infectivity was age-dependent in relation to salinity. Larvae were infective to intermediate and paratenic hosts for up to 80 d in fresh water, 21 d in 50% sea water and up to 8 d in 100% sea water. The data confirm field observations that infection levels decrease with an increase in salinity. The study contributes to experimental verification of the colonization abilities of A. crassus and supports the hypothesis that A. crassus can be disseminated and transmitted in brackish water. The importance of regular monitoring and stringent hygiene practices in the transportation of eels is emphasized.

Survival and transmission of Anguillicola crassus Kuwahara, Niimi & Itagaki, 1974 (Nematoda) in seawater eels.

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.

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.

Impact of low water temperature on the development of Anguillicola crassus in the final host Anguilla anguilla.

The effect of low water temperatures on the development and viability of larval and adult Anguillicola crassus (Nematoda) in the final host Anguilla anguilla was studied. European eels were experimentally infected with A. crassus and then maintained for 4 mo at 4, 9, 10, and 19 degrees C. Larval development showed a temperature-dependent pattern and was significantly retarded at low temperatures. Third-stage larvae survived a 4 mo period at 4 degrees C without being affected, although they were not able to invade the swimbladder wall at this temperature. In contrast, adult worms were severely harmed during a 4 mo period at 4 degrees C, as reflected by increased mortality and decreased growth and reproductivity as compared to the worms maintained for the same period at 18 degrees C. Starvation for the eels at 4 mo at 19 degrees C did not affect the development and growth of the nematode. The experimentally obtained results support the hypothesis that the spread of A. crassus in boreal regions, e.g. Northern Europe, is restricted by the natural ambient temperature regimes.

Distribution and prevalence of Anguillicola crassus (Nematoda) in eels Anguilla anguilla of the rivers Rhine and Naab, Germany.

Over a period of 2 yr eels were collected at several sites along the river Rhine as well as from adjacent waters and examined for Anguillicola crassus in the swimbladder. Additionally eels were collected once from a site on the river Naab (Danube system). All eels were searched for both juvenile and adult stages of A. crassus, revealing prevalences of 80% in eels from the river Rhine and 39% in eels from the river Naab. The mean intensities were 5.6 and 7.7, respectively. With respect to the prevalence and mean intensity of infection, no significant differences were observed between the different sampling sites in the Rhine system. Neither the prevalence nor the intensity showed significant seasonal patterns. Further, no correlation between the condition factor of the eels and intensity of infection with A. crassus could be found. Of all eels examined, 28% showed pathological alterations of the swimbladder.

Attempts to analyse Anguillicola crassus infection and the humoral host response in eels (Anguilla anguilla) of Lake Balaton, Hungary.

Since the introduction of Anguillicola crassus into Europe, anguillicolosis has been a considerable problem in several countries. From 1991, periodical eel mortality occurred in Lake Balaton, Hungary. However, eels with a worm burden of 20 to 50 parasites did not show severe swimbladder lesions, which observation cast doubts on the primary aetiological role of the parasite in the eel kill. In order to study the pathology of the infection, from the spring of 1996 until October of the same year, 51 eels were collected from two regions of Lake Balaton and examined for swimbladder changes. To detect humoral antibodies, an enzyme-linked immunosorbent assay (ELISA) was performed, using cuticular-oesophageal worm antigen. The results of the test show the applicability of the method. However, no direct correlation was found between antibody levels or the intensity of infection and the swimbladder lesions. The low level of specific antibodies and the increasing severity of swimbladder changes in the autumn suggest that parasite-induced immunity is insufficient to prevent reinfection.

Present occurrence of Anguillicola novaezelandiae (Nematoda: Dracunculoidea) in Europe and its development in the intermediate host.

An examination of a sample of European eels, Anguilla anguilla (L.), collected from Lake Bracciano near Rome in 1993, the only known European locality with the occurrence of the introduced swimbladder nematode Anguillicola novaezelandiae Moravec et Taraschewski, 1988, revealed for the first time the presence of two Anguillicola species, A. novaezelandiae and A. crassus. In view of the investigations carried out by current authors in Bracciano Lake in the years 1982-1992, it is apparent that the latter species has been introduced into the lake quite recently, where it quickly became a dominant species. The development of A. novaezelandiae was experimentally studied in the copepod intermediate host, Cyclops strenuus, for the first time. The copepods were infected with nematode second-stage larvae at 21-22 degrees C; fully developed infective third-stage larvae were obtained 13 days p.i. The general morphology of individual larval stages of A. novaezelandiae was similar to that of larvae of the related species A. crassus.

Third-stage larvae of Daniconema anguillae (Nematoda: Dracunculoidea) in the subcutaneous tissue of eel Anguilla anguilla.

Daniconema anguillae Moravec et Køie, 1987 larvae measuring 1.64-1.76 mm were occasionally found in considerable numbers in the fins and subcutaneous connective tissue of approximately 50% of eel Anguilla anguilla (L.) sampled from Lake Balaton, Hungary. The larvae were noted for their slender body, very long tail with a rounded tip, a densely transversely striated cuticle, and the presence of boring tooth and large kidney-shaped amphids on the cephalic end. The larvae could easily be recovered from the above mentioned organs by placing them into isotonic saline solution. No disease signs or pathological changes attributable to the larval infection could be observed. The only histological indication of host reaction was the appearance of macrophages adhering to the body surface of larvae and of cells with spherical nucleus in areas around the larvae. A possible life cycle pattern of D. anguillae is discussed.