Phenotypic modification of roach (Rutilus rutilus L.) infected with Ligula intestinalis L. (Cestoda: Pseudophyllidea).

In European freshwater, cyprinid fish may be heavily infected by plerocercoids of the pseudophyllidea cestode Ligula intestinalis (L.). During their development, these parasites grow rapidly to a large size in the fish's body cavity, characteristically distending the abdomen. In this study, the influence of this tapeworm on roach (Rutilus rutilus L.) morphology was analyzed. Forty-five infected and 45 uninfected roach were collected from the Lavernose-Lacasse gravel pit in Toulouse, south western France and examined for 40 morphological measurements to study phenotypic modification of the body and 14 bilateral characters for an analysis of asymmetry. Results indicate that the degree of bilateral asymmetry does not change between infected and uninfected roach, despite the strong host-morphological modifications such as deformation of the abdomen, fin displacements at the level of the tail, and sagging of the vertebral column. The intensity of abdominal distension and fish morphology changes depends on the total parasite biomass present. Differences were observed in morphology at different levels of infection, which relate to established effects of L. intestinalis on the physiology and behavior of intermediate hosts. These morphological changes induced by the parasite could increase trophic transmission to the definitive avian hosts.     

Behaviour of roach (Rutilus rutilus L.) altered by Ligula intestinalis (Cestoda: Pseudophyllidea): a field demonstration

1. We studied the influence of a cestode parasite, the tapeworm Ligula intestinalis (L.) on roach ( Rutilus rutilus L.) spatial occupancy in a French reservoir (Lake Pareloup, South-west of France).
2. Fish host age, habitat use and parasite occurrence and abundance were determined during a 1 year cycle using monthly gill-net catches. Multivariate analysis [generalized linear models (GLIM)], revealed significant relationships ( P  < 0.05) between roach age, its spatial occupancy and parasite occurrence and abundance.
3. Three-year-old roach were found to be heavily parasitized and their location toward the bank was significantly linked to parasite occurrence and abundance. Parasitized fish, considering both parasite occurrence and abundance, tended to occur close to the bank between July and December. On the contrary, between January and June no significant relationship was found.
4. These behavioural changes induced by the parasite may increase piscivorous bird encounter rate and predation efficiency on parasitized roach and therefore facilitate completion of the parasite's life cycle.     

The effects of a change in the growth rate of roach, Rutilus rutilus (L.), on the biology of the fish tapeworm Ligula intestinalis (L.)

Investigations into the biology of the roach and the pathogenic tapeworm Ligula intestinalis (L.) populations at Slapton Ley, Devon were carried out between October 1982 and December 1984. Data collected from the lake since 1977 have also been re-analysed to determine how the dramatic improvement in the individual growth rate of the roach over this period may have affected the growth, maturation and life-cycle of Ligula. Only the very young roach at this site become infected, so it was possible to follow cohorts of plerocercoids of similar age through each roach year class. Results for the 1978 and 1983 year classes are presented in detail. The roach grew extremely rapidly from May to August in each year, resulting in a pronounced cyclical pattern of changes in the condition of the roach, with the lowest condition occurring in late winter and spring. The yearly increase in the roach growth rate was accompanied by an increase in plerocercoid growth rate in the 0 + roach, but not in the 1 + roach. The growth rate of the plerocercoids was very high compared to that at other sites. It is usual for the parasite index (PI) of Ligula-infected fish to be high and to increase throughout their first few years of life. At Slapton, however, a lack of multiple infections has prevented high PIs from occurring, and in recent years the growth rate of the roach has been so high that the plerocercoids were unable to maintain a high weight relative to the fish, and the highest PIs occurred in the 0 + roach throughout late winter and spring. In recent year classes, therefore, the maximum PIs and highest pathogenicities coincided with the period of lowest condition in the 0+ roach. Observations of both caged and natural populations of 0+ roach over winter showed that a significant loss of roach containing the larger plerocercoids occurred from the population. In vitro cultivation of Ligula plerocercoids showed that they were capable of maturation at weights of 0.5 g, and only 6 months after having infected the roach. The increase of the growth rate of the plerocercoids in the 0+ roach has therefore resulted in a greater proportion of these plerocercoids being capable of infecting the definitive host. As a result of the increase in individual growth rate of the roach at Slapton, the potential for Ligula transmission, as measured in terms of both their pathogenicity and maturity, has shifted from the 1 + to the 0+ roach.     

Peculiarities of lipid peroxidation indices in Ligula intestinalis (Cestoda: Pseudophyllidea) and its host--Abramis brama (L.)

A comparative analysis of indices of peroxidation lipids in tissues of Ligula inteslinalis plerocercoids and in the intermediate fish host, the bream Abramis brama, was performed for the content of common lipids (CL), malonate di-aldehyde (MDA), which is a product of lipid peroxidation (POL), and common antioxidate activity (CAA). The dependence of indices upon size of parasites is recovered. The long-sized parasites had higher rate of MDA and intense CAA. The higher rate of MDA and low one of CAA was observed in the infected fishers comparing to the healthy ones.     

Inhibition of gametogenesis by the cestode Ligula intestinalis in roach (Rutilus rutilus) is attenuated under laboratory conditions

Reproductive parameters of Ligula intestinalis-infected roach (Rutilus rutilus) which were held under long-tem laboratory conditions with unlimited food supply were investigated. Although uninfected and infected roach showed no difference in condition factor and both groups deposited perivisceral fat, the gonadosomatic-index was significantly lower in infected female and male roach. Quantitative histological analysis revealed that gonad development was retarded upon parasitization in both genders. In contrast to the phenotype described in the field, infected females were able to recruit follicles into secondary growth, but a high percentage of secondary growth follicles underwent atresia. In both genders, the histological data corresponded well with reduced expression of pituitary gonadotropins and lowered plasma concentrations of sex steroids, as revealed by real-time RT-PCR and ELISA, respectively. Furthermore, a reduction of vitellogenin mRNA and modulated expression of sex steroid receptors in the liver was demonstrated. Like in the field, there was a significant adverse impact of L. intestinalis on host reproductive physiology which could not be related to parasite burden. Our results show, for the first time, that maintenance under laboratory conditions can not abolish the deleterious effect of L. intestinalis on gametogenesis in roach, and indicate a specific inhibition of host reproduction by endocrine disruption.     

Ontogenetic changes in haemoglobins in roach, Rutilus rutilus (L.) and rudd, Scardinius erythrophthalmus (L.)

The haemoglobins and globins of the roach, Rutilus rutilus (L.) and the rudd, Scardinius erythrophthalmus (L.) have been studied by starch gel electrophoresis. At all stages of development studied, the roach and rudd haemoglobins give a similar electrophoretic pattern, the young fish of about 2 cm standard length possessing three separate haemoglobins, of which two bands only persist in the adult. The evolutionary significance of multiple haemoglobins in fishes is briefly discussed.     

Isolation and characterization of microsatellite loci in the tapeworm Ligula intestinalis (Cestoda: Pseudophyllidea)

Ligula intestinalis is a species with a complex biology. Several strains, distinct in the host specificity and biogeography, are supposed to exist; however, their morphological or molecular identification remains unresolved. We describe the isolation of 16 polymorphic microsatellite markers. The number of alleles ranged from four to 14 in a population of 21 L. intestinalis individuals. Loci showed amplification success across samples of worldwide distribution. A cross‐amplification of 14 markers in Digramma interrupta (Cestoda: Pseudophyllidea) was observed.     

Fertility in first-generation hybrids of roach, Rutilus rutilus (L.), and silver bream, Blicca bjoerkna (L.)

Fertility in first‐generation hybrids of roach, Rutilus rutilus, and silver bream, Blicca bjoerkna, was investigated. Sperm and egg production of hybrids at first sexual maturity were examined. Eggs from female hybrids were artificially fertilized with the sperm of a corresponding hybrid male; a hybrid male from the reciprocal crossbreeding; a parental species male R. rutilus; and a parental species male B. bjoerkna. The results revealed that gametogenesis was normal in female hybrids. However, in male hybrids, a low efficiency of gametogenesis was observed. The semen of male hybrids was extremely dilute, with spermatozoa concentration lower than that in parental species. Nevertheless, these F1 hybrids (males and females) from reciprocal crossbreeding were fertile. F2 and backcross generations were produced, but F2 crosses from the female hybrid and corresponding hybrid male displayed a drastically slower hatching rate. Also higher proportions of deformed embryos were hatched than in other post‐F1‐generation crosses.     

Feeding migration of roach, Rutilus rutilus (L.), in Lake Arungen, Norway

In the period April–July 1980 we studied the feeding migration and food of roach in a small tributary of the eutrophic Lake Årungen, south-eastern Norway. Tagging experiments revealed a tendency in roach to utilize a specific tributary both for feeding and for spawning. The mean size of roach ascending the tributary in late June and July was significantly larger than the mean size of roach spawning 1–2 months earlier, probably due to higher water discharge in July than in May and June. The experiment indicates two separately motivated migrations involving homing. The roach fed more profitably in the tributary than in the lake, both in terms of food availability and predator avoidance.     

Some aspects of the biology of gudgeon Gobio gobio (L.) in Irish waters

Data on some aspects of the biology of gudgeon were obtained during a programme of research on Irish coarse fishes. Eggs of gudgeon, were collected on 3 occasions—on June 8 and again on June 29, 1965 in a small rivulet entering a hydro-electric reservoir in Co. Cork; and on June 2, 1966 in a millstream on the Grand Canal in Co. Dublin. The eggs were greyish-white in colour, opaque, and 1·375–1·625 mm diam. Newly-hatched larvae from eggs from Co. Cork measured 5·45 mm and were distinguished by the presence of black pigment largely confined to the ventral aspect of the yolk sac and tail. They began to feed after 3 days; in 5–6 days the yolk was fully absorbed, the fish then measuring 6·2 mm. Following the creation of 2 hydro-electric reservoirs on the River Lee in Co. Cork, large populations of gudgeon developed in them for a time. It was found that, in the reservoirs, gudgeon less than 7·5 cm were feeding mainly on Cladocera, especially Chydoridae. The larger reservoir gudgeon and gudgeon in rivers and streams in Co. Cork fed mainly on molluscs, Gammarus, chironomid larvae, caddis larvae and ephemeropteran nymphs. Some gudgeon held a good deal of filamentous algae. Gudgeon were aged by means of scale reading, the results of which were confirmed by length-frequency distribution analysis in the case of the reservoirs. In the Lee system, gudgeon grew relatively rapidly at first, attaining a mean fork-length at 3 years old of 9·5–10·8 cm according to year-class. In the River Allow, a Cork Blackwater tributary, early growth was slower than in the Lee, but the length increments in the third and subsequent seasons were bigger than in the Lee reservoirs. In the Lee, most gudgeon of both sexes spawned for the first time when 2 years old, at a minimum fork-length of 6·9 cm in the case of males and 7·9 cm in the case of females. The trematode parasite Paracoenogonimus ovatus was of frequent occurrence in Irish gudgeon aged 2 years and upwards.     

Histochemical characterization of myotomal muscle in the roach, Rutilus rutilus (L.)

A histochemical study of the myotomal muscles in the roach revealed three main muscle regions: red, intermediate and white. These were distinguished on the basis of glycogen content, succinate dehydrogenase (SDH), and myofibrillar ATPase (mATPase) activity. Except for the red fibre region, none of these described regions is homogeneous. The principal new findings are the toniclike fibre, the presence of a transitional zone with two fibre types, and the mosaic organization of the white fibre region. The significance of this type of myotome architecture in relation to the locomotion of the species is discussed.     

A search for introgressive hybridization in the rudd, Scardinius erythrophthalmus (L.), and the roach, Rutilus rutilus (L.)

Rudd, roach and their hybrids from an interbreeding population at Norwich were examined electrophoretically for seven enzymes; an examination of pharyngeal teeth supplemented the data. The rudd were compared to rudd from Lake Trawsfynydd, Wales, a lake free from roach. No differences were observed between the rudd from the two localities and no evidence was found of introgression within the fish from Norwich.     

The morphology and histology of the pineal organ in roach, Rutilus rutilus (L.)

The pineal organ in the roach, Rutilus rutilus (L.), is covered by a semi-transparent area, the pineal window. Beneath this the pineal is attached to a long robust stalk, lying just under the parietal bone. The pineal is attached to the brain through the dorsal sac. Three cell types have been identified histologically. These are the sensory cells, supporting cells and the ganglia cells. The inner segment of the sensory cells respond to PAS and AF staining, while the remaining cells respond to Orange G, LG, or Acid Fuchsin. The evidence suggests that the roach pineal may have a dual photosensory and glandular function.     

Electrophoretic identification of roach (Rutilus rutilus L.), rudd (Scardinim erythrophthalmus L.), bream (Abramis brama L.) and their natural hybrids

Roach, rudd, bream and their natural hybrids of 2 cm standard length or larger can be definitively identified by their enzyme electrophoretic patterns. Zymograms of lactate dehydrogenase and esterases as produced by vertical starch gel electrophoretic analysis of whole fry or adult eye extracts are the most useful in this respect. The lactate dehydrogenase isozymes, containing B sub-units, migrate more anodally in rudd and bream than in roach. Due to the tetrameric structure of lactate dehydrogenase, in hybrid rudd x roach and roach x bream, eleven isozymes can be observed as compared with six in the parental patterns. Esterases show unique patterns for all species and hybrids. With the exception of one fraction in rudd x bream, the esterase patterns of hybrids show summations of the parental phenotypes.     

Sperm quality, secondary sexual characters and parasitism in roach (Rutilus rutilus L.)

According to sperm competition models, a male spawning in a disfavoured role should have spermatozoa with higher velocity but shorter longevity compared with a male spawning in a favoured role. Moreover, immunosuppressive androgens are needed to produce both secondary sexual characters and sperm cells. The 'sperm protection' hypothesis suggests that the immunosuppressive action of androgens has evolved to protect haploid spermatozoa, which are antigenic, from autoimmune attacks. Therefore, a male with high sexual ornamentation may be more susceptible to diseases but may possess better quality ejaculate than his less ornamented rival. We studied sexual ornamentation (breeding tubercles), ejaculation quality (sperm concentration, longevity and spermatozoal velocity) and intensity of parasitism in the cyprinid, Rutilus rutilus . Sperm longevity and spermatozoal velocity were positively correlated. Males having elaborated sexual ornamentation had longer-lived sperm and more Myxobolus mülleri parasites in the liver compared with males with low ornamentation. However, no difference was found between males with different degrees of ornamentation with respect to sperm concentration, spermatozoal velocity or other parasites. Since the highly ornamented males had higher sperm longevity, the present results are partly consistent with the predictions of the sperm competition models and the 'sperm protection' hypothesis.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 81 , 111–117.     

The induced spawning of roach, Rutilus rutilus (L.), with the antioestrogens clomiphene and tamoxifen

The induced spawning of gravid roach was investigated using the antioestrogens clomiphene and tamoxifen. Three dose levels ofeach were used: 0·1, l or 10 mgkg^-1, given twice with a 4-day interval to groups of eight fish with saline controls. Running males were randomly distributed. Tamoxifen, when injected at a rate of l mg kg^-1 was found to be most successful. This treatment induced ovulation in five of the six females, and profuse spawning on 3 consecutive days, 4 days after the first injection. Clomiphene induced ovulation and spawning in one of six females at 2 × 10 mg kg^-1, and two of eight females at 2 × l mg kg^-1, respectively 6 and 7 days after the first injection. The eggs produced showed normal devel-opment. No control fish ovulated or spawned. Both drugs probably act by indirect mechanisms, blocking sex steroid feedback inhibition of gonadotropin (GtH) secretion at the pituitary, thereby inducing a plasma GtH surge. The results of this experiment suggest that tamoxifen may be an effective substitute for pituitary preparations in the induced spawning of fish.     

Comparative cytogenetics of Diphyllobothrium ditremum (Creplin, 1925) and Ligula intestinalis (Linnaeus, 1758) (Cestoda: Pseudophyllidea)

An investigation carried out on two species of pseudophyllidean cestodes belonging to different families showed very close karyological affinity between them. The karyotypes of Diphyllobothrium ditremum and Ligula intestinalis both consist of 18 bi-armed chromosomes and are almost identical with respect to the relative length and the centromeric indices of corresponding chromosomes. Statistically significant differences exist in the morphology of chromosomes 2 and 4, but they are not striking and may be due in part to errors of measurement. Differences in the absolute length of the chromosomes were noted: the chromosomes of D. ditremum are somewhat larger (from 2.7 to 8.5 μm) than those of L. intestinalis (from 1.9 to 5.4 μm). The results obtained were compared with data existing for other pseudophyllidean cestodes and preliminary conclusions on the karyotypic evolution in that group of helminths were made.