Effect of domestication on ide Leuciscus idus L. reproductive parameters

Stock enhancement is the main source of Acipenser gueldenstaedtii and A. stellatus recruitment in the Sea of Azov, where up to 30 million juveniles have been released in recent years. The hatchery production strategy aimed solely to produce and release the largest possible number of juveniles, utilizing only the most mature broodfish of the spring spawning run. Although monitoring of age, mass and physiological conditions of the broodstock during the period of hatchery breeding did not show significant changes, the spawning run was dramatically shortened from several months to just 15 days, and includes only fish with advanced gonadal maturity. Conservation of the Azov Sea species and their unique spawning ecotypes is currently supported by the Federal Living Gene Bank, which maintains over 9000 adults of eight endangered species. This paper summarizes the results of comparative analysis of biological characteristics such as growth, age of the first sexual maturation, relative fecundity, and morphological and physiological indices in wild and cultured specimens of different species and intraspecific groups of sturgeons. We conclude that there is a need for developing better hatchery technologies to maintain diversity of the stocks, through breeding protocols that maximize the genetic diversity of offspring based on evaluation of parentage and relatedness in farmed stocks by microsatellite loci.     

Early development and allometric growth patterns in ide Leuciscus idus (Linnaeus 1758)

Allometric growth and ontogeny were studied in ide Leuciscus idus (L.) reared in laboratory conditions from 0 to 30 days post hatching (DPH), to define important steps in the species’ early life history. Based on the external morphology, eleven different stages during the early development of ide were identified (ES9_a – JS1_a; ES – embryonic stage, JS – juvenile stage). After a short period of rest, hatched individuals were visual feeders with a pelagic life. Organogenesis and allometric changes indicated that development priorities concerned feeding efficiency, respiration efficiency and locomotion performances. Also concluded was that the gradual changes observed in body proportions (which occurred across a wide range of body lengths) reflect the absence of abrupt changes in the behaviour and habitat and may also be considered as a gradual adaptation to life in flowing water.     

Influence of rearing method on ide Leuciscus idus L. juvenile survival under pike and pikeperch predation

Nase Chondrostoma nasus L., once one of the most common fish species in the upper reaches of central European rivers, is now considered endangered throughout its natural range. Since 2000 the Department of Ichthyobiology and Fisheries, Warsaw Agricultural University, has co-operated with the Polish Anglers Association in Krosno to study the effects of river restocking with pond‐farmed nase juveniles. The experiment is carried out on Wislok River, where nase was very twenty years ago but has now completely disappeared. Selected parts of the river are stocked with tagged, one‐summer and 1 year‐old juveniles. Changes in nase occurrence, and their growth and foraging behaviour have been analysed.     

Preliminary characterization of a reovirus isolated from golden ide Leuciscus idus melanotus

Some characteristics of a reovirus recently isolated from golden ide Leuciscus idus melanotus and tentatively designated as golden ide reovirus (GIRV) were determined. Spherical non-enveloped particles with an outer capsid of about 70 nm and an inner capsid of about 50 nm were observed by electron microscopy. The density of the virus determined in CsCl gradients was 1.36 g ml-1. The genome contained 11 segments of dsRNA. GIRV differed from other aquareoviruses by a slight reduction of infectivity after treatment with chloroform and by the absence of forming syncytia in cell monolayers.     

Viral diseases of fish: first report of carp pox in golden ide (Leuciscus idus) in North America

Carp pox, a putative viral disease exotic to North America, occurred in golden ide 1 yr after the fish were imported into the United States from the Federal Republic of Germany. The raised, white, plaque-like lesions, which occurred on about 5% of the fish, healed spontaneously and caused no mortality. Electron micrographs showed herpesvirus-like particles associated with lesion specimens; however, no infectious viruses were detected in tests with seven warmwater fish cell lines.     

Identification and characterization of microsatellite DNA markers developed in ide Leuciscus idus and Siberian roach Rutilus rutilus

The first five microsatellite markers for the ide, Leuciscus idus, and four microsatellite markers for the Siberian roach, Rutilus rutilus, were designed. Cross‐amplification of ide markers was examined in Siberian roach and vice versa. The number of alleles per locus ranged from three to 13 in ide and from two to eight in roach. The expected heterozygosity ranged from 0.313 to 0.909 in ide and from 0.119 to 0.775 in roach. These markers could be used to evaluate the genetic population structure of these species and other fish from the Cyprinidae family.     

The modular habitat model (MHM) for the ide, Leuciscus idus (L.) - a new method to predict the suitability of inshore habitats for fish

Groynes are the dominant river engineering structures along the lowland section of large European rivers such as the Rhine, Danube and the Elbe. More than 6000 groynes structure the 400 km stretch of the potamal of the Elbe River. After 1945, destruction of the groynes increased through ice and flood events in the eastern part of Germany. In the past ten years, groyne reconstruction was accompanied by a controversial discussion in the context of the ecological integrity of the Elbe River. With the modular habitat model (MHM) a tool was developed to evaluate the suitability and to balance the availability of fish habitats in groyne fields of different conditions. The morphodynamic module produced a digital terrain model and a spatial model of flow velocity for each groyne field separately. Based on point abundance sampling by electro‐fishing, models of habitat preference were developed for different life history stages by logistic regression. Statistical models predicting the preference of fish‐environment relationships (Leuciscus idus) at different life history stages. The models were discriminated and validated by receiver operating characteristic (ROC) curves. The link between the statistical and the spatial model was realised in the suitability module. The suitability of microhabitats is expressed in classes for each species and life history stage separately. Habitat availability is balanced on the level of mesohabitat, e.g. different types of groyne fields. The temporal dynamics of habitat availability are analysed by considering different levels of discharge. For the stage ‘juvenile A’ and preadult the habitat suitability is better in fields downstream of destroyed groynes. For ‘juvenile B’ and adult stages of the ide, groyne fields in general constitute low habitat suitability. Differences in spatial availability are higher than the differences in temporal habitat availability.     

Light- and electron-microscope studies on the pseudobranch of the golden orfe, Leuciscus idus L.

The structure of the pseudobranch of the golden orfe is described from light- and electron-microscopical observations. It is characterized as a covered pseudobranch overlaid by a mucous membrane. The pseudobranch of the golden orfe has no respiratory function, but the electronmicroscopical results indicate that it could function endocrinally by registering and regulating salinity or osmolarity. The epithelial cells are represented by pseudobranch-type cells. No chloride cells were seen. Rodlet cells are located near blood vessels at the base of the pseudobranch and in the covering epithelial layer, together with mucous cells and mast cells. In the endothelial intima of the arteries the so-called Weibel-Palade bodies are described.     

Genetic variation and divergence of Leuciscus pyrenaicus and L. carolitertii (Pisces, Cyprinidae)

The taxonomy of the genus Leuciscus in the Iberian Peninsula has been presented on the basis of morphological analysis. Two species are believed to occur in Portuguese inland waters, L. carolitertii , the northern basins chub, and L. pyrenaicus , which was described for the Tejo and southern basins. Since only slight differences in the meristic and morphometric characters are observed, the distinction between the two species has been made previously mainly on the basis of osteology. In order to investigate the patterns of genetic differentiation between these species, 24 sites were sampled for both species, and examined electrophoretically for the study of variation at 27 presumptive loci. The results pointed to different levels of population differentiation among drainages and support the recognition of the species level for L. pyrenaicus and L. carolitertii. Our results also suggested that the distribution area of L. pyrenaicus is probably restricted to Tejo and to the small drainages near Tejo, while L. carolitertii seems to have a widespread distribution area, including Lima, Douro, Vouga, Mondego, Guadina and Sado drainages. Moreover, concerning the southern populations, Mira and Arade, the surprisingly high level of differentiation observed at several loci, supports the existence of a further species occurring in these small drainages. The evolutionary aspects related to that differentiation are discussed.     

Mass marking of Leuciscus idus larvae using Artemia salina as a vector of fluorescent dyes

A method for the mass marking of ide Leuciscus idus larvae by feeding them Artemia salina nauplii that were immersed in different solutions of alizarin red S, tetracycline hydrochloride and calcein was tested. The best quality marks were obtained after feeding fish for 4 days with nauplii that had been immersed in 200 mg l^?1 alizarin red S.     

Purified scrapie prions resist inactivation by UV irradiation.

The development of effective purification protocols has permitted evaluation of the resistance of isolated scrapie prions to inactivation by UV irradiation at 254 nm. Prions were irradiated on ice with doses of UV light ranging up to 120,000 J/m2. UV dosimetry experiments, performed with Saccharomyces cerevisiae plasmid DNA or eucaryotic cells, indicated that under these experimental conditions an incident UV dose of 10 J/m2 formed 2 thymine dimers per 5.1 X 10(6) daltons of eucaryotic cell DNA. The D37 values for scrapie prions ranged from 17,000 to 22,000 J/m2; D37 values were also determined for virus, viroid, and enzyme controls. The number of pyrimidine dimers formed was correlated with the D37 values obtained for irradiated prions and target nucleic acids. The D37 value for bacteriophage M13, 6.5 J/m2, occurred at a dose that would form 0.56 dimers per target genome; the D37 for potato spindle tuber viroid, 4,800 J/m2, occurred at a dose that would form about 24 dimers per target viroid. The D37 value for an EcoRI restriction site, a target of 12 bases, occurred at a dose that would correspond to the formation of 0.89 thymine dimers per target site. The D37 value for prions occurred at a dose that would form 1 dimer in every 4 bases of single-stranded target nucleic acid. If the putative scrapie nucleic acid were double-stranded and readily repairable after UV damage, then the prion D37 value could reflect a nucleic acid molecule of 30 to 45 base pairs. While the D37 value for prions fell within the range of pure protein targets, our experiments cannot eliminate the possibility that a prion contains a small, highly protected nucleic acid molecule.     

Spironucleus vortens (Diplomonadida) in the Ide,Leuciscus idus (L.) (Cyprinidae): a warm water hexamitid flagellate found in northern Europe

The hexamitid flagellate Spironucleus vortens, previously reported from Pterophyllum scalare from Florida, was found in the intestine of Leuciscus idus in Norway. The flagellate was cultivated and studied by scanning and transmission electron microscopy. Identification was based on a suite of ultrastructural features unique for S. vortens: compound lateral ridges, a swirled posterior end, and a distinctive microtubular cytoskeleton. Microfibrillar structures with a periodicity of 0.13 microm in the right peripheral part of the compound lateral ridges were shown to be responsible for the distinctive rope-like appearance of the peripheral ridge seen in scanning electron micrographs, and not previously reported for S. vortens. The present results show a wide geographic distribution and a wide temperature tolerance for S. vortens. The flagellate was successfully cultivated at 5 degrees C and 15 degrees C, having previously been cultivated between 2-34 degrees C. Spironucleus vortens is believed to be endemic in Norwegian waters, but an introduction hypothesis is also discussed. The similarity is striking between S. vortens and S. elegans, previously described from amphibians and fish in Europe, and the possibility of conspecificity is believed to be high.     

Increased inactivation of Saccharomyces cerevisiae by protraction of UV irradiation.

The principle of equi-effectivity of the product of intensity and exposure time (principle of Bunsen-Roscoe) of UV irradiation has been assumed to be valid for the inactivation of microorganisms in general. Earlier studies claimed higher survival of Escherichia coli B/r with fractionated irradiation compared with single-exposure survival. However, data on the inactivation effect of protraction of UV irradiation are not available. By means of a specially designed UV irradiation apparatus which secured absolute UV dose measurements throughout the experiments, the effects of variation of UV irradiation intensities (253.7 nm) and exposure times were tested on the inactivation of a bacterial virus (Staphylococcus aureus phage A994), a vegetative bacterial strain (E. coli ATCC 25922), and bacterial spores (Bacillus subtilis ATCC 6633) as well as three haploid laboratory strains (RC43a, YNN281, and YNN282) and two diploid strains (commercial bakery yeast strain and laboratory strain YNN281 x YNN282) or yeast (Saccharomyces cerevisiae) and spores of the latter diploid yeast strain. Each test organism was exposed to three UV intensities (0.02, 0.2, and 2 W/m2), with corresponding exposure times resulting in three dose levels for each intensity. Differences in inactivation rates were tested by analyses of variance and Newman-Keuls tests. Virus and bacteria showed no differences in inactivation rates by variation of intensities and exposure times within selected UV doses; hence, the principle of Bunsen-Roscoe could not be rejected for these strains. However, in the eukaryotic test strains of S. cerevisiae longer exposure times with lower intensities led to enhanced inactivation in both haploid and diploid strains, with a more pronounced effect in the diploid yeast strains, whereas in yeast spores in this dose rate effect could not be observed.     

Genetic variability and differentiation in Leuciscus (Telestes) souffia. Taxonomic and conservation inferences

Genetic variability of eight populations of Leuciscus (Telestes) souffia from France, Italy and Slovenia, and one of Leuciscus (Squalius) carolitertii from Spain was analysed using allozymic electrophoresis. For L (Telestes) souffia the parameters of variability ranged from P = 1 8.2 to 48.5 % and H_o = 0.023 to 0.092, values included in the global averages found for freshwater fishes. The phylogenetic relationships among these populations lead us to consider the souffia species belonging to the genus Telestes, and to subdivide it into two species: souffia (with at least two subspecies: souffia in France and agassizi in Slovenia), and muticellus. The genetic structure found within the French populations showed a certain differentiation between the Hérault Basin and the Var/Argens populations, recommending the preservation of these entities that could represent different adapted genetic pools. In the same way, the Italian region should be studied in detail, since the populations analysed from Bevera and Staffora Rivers (T. muticellus) showed evidence of a break of genetic flow (including a diagnostic locus between them).     

Genetic individualization of sable (Martes zibellina L. 1758) using microsatellites

Genetic individualization based on non-invasive sampling is crucial for estimating the numbers of individuals in endangered mammalian populations. In sable (Martes zibellina)-poaching cases, identifying the number of animals involved is critical for determining the penalty. In addition, investigating animal numbers for wild sable populations requires genetic individualization when collecting several samples in neighboring regions. Microsatellites have been demonstrated to be reliable markers for individual identification. Thirty-three microsatellite loci derived from Mustelidae were selected to develop a genetic individualization method for sable. Three reference populations containing 54 unrelated sables were used to calculate allele number, allelic frequencies, and the polymorphic information content of each locus. The data were subsequently used to assess the validity of a combination of twelve loci for sable individualization. We defined twelve polymorphic loci that were easy to be amplified and genotyped. Four significant deviations from Hardy-Weinberg equilibrium were observed among the 12?loci in the three populations. The match probability of an individual from the reference populations with a random individual based on the 12?loci was 1.37?×?10^?13. Using the combination of the twelve loci provides sufficient power to individualize sables considering the levels of microsatellite polymorphism observed. These loci were successfully applied to a case of sable poaching and provided valid evidence to determine the penalty. The genetic individualization of sable based on these loci might also be useful to investigate the numbers of animals in wild populations.     

A comparative study of neuropeptides in the intestine of two stomachless teleosts (Poecilia reticulata,Leuciscus idus melanotus) under conditions of feeding and starvation

Summary Endocrine cells containing bombesin-, enkephalin-, gastrin/CCK-, 5-HT-, and substance P-like material were demonstrated in the alimentary tract of Poecilia reticulata and Leuciscus idus melanotus. Endocrine cells with neuropeptide-Y-like immunoreactivity were found only in P. reticulata, those with VIP-like immunoreactivity only in L. idus melanotus. Gut nerves showing bombesin-, G/CCK-5-HT-, neurotensin-, substance P-and VIP-like immunoreactivity were observed in both species investigated, enkephalin- and neuropeptide Y-like immunoreactivity in P. reticulata alone. The distribution and amount of endocrine cells and nerves along the gut as visualized with the appropriate antisera varied in both teleosts. Histologically, the intestinal tract of these stomachless fish can be divided into three regions. A large number of endocrine cells with VIP-like immunoreactivity was noted in the rectum of L. idus melanotus. Endocrine cells containing bombesin-, enkepha-lin- and substance P-like material were found only in intestinal parts I and II in L. idus melanotus. Neuropeptide Y-like immunoreactivity was absent from intestinal part I of P. reticulata. The influence of starvation on the immunoreactivity of nerves and enteroendocrine cells in the teleost intestine was examined. After a starvation period of more than 6 weeks, no alterations were observed either in the appearance or amount of nerve and endocrine cell immunoreactivity.     

Genetic control of mitotic crossing-over in yeasts. III. Induction by 8-methoxypsoralen and long-wave UV irradiation (lambda=365 nm)

The lethal effect of 8-methoxypsoralen (8-MOP) plus 365 nm light has been studied in haploid radiosensitive strains of Saccharomyces cerevisiae. The diploid of wild type and the diploid homozygous for the rad2 mutation (this mutation blocks the excision of UV-induced pyrimidine dimers) were more resistant to the lethal effect of 8-MOP plus 365 nm light than the haploid of wild type and rad2 haploid, respectively. The diploid homozygous for rad54 mutation (the mutation blocks the repair of double-strand breaks in DNA) was more sensitive than haploid rad54. The method of repeated irradiation allowed to study the capacity of radiosensitive diploids to remove monoadducts induced by 8-MOP in DNA. This process was very effective in diploids of wild type and in the rad54 rad54 diploid, while the rad2 rad2 diploid was characterized by nearly complete absence of monoadduct excision. The study of mitotic crossing over and mitotic segregation in yeast diploids, containing a pair of complementing alleles of the ade2 gene (red/pink) has shown a very high recombinogenic effect of 8-MOP plus 365 nm light. The rad2 mutation slightly increased the frequency of mitotic segregation and mitotic crossing over. The rad54 mutation decreased the frequency of mitotic segregation and entirely suppressed mitotic crossing over. The method of repeated irradiation showed that the cross-links, but not monoadducts, are the main cause of high recombinogenic effect of 8-MOP plus 365 nm light. The possible participation of different repair systems in recombinational processes induced by 8-MOP in yeast cells is discussed.     

Larval rearing of chub, Leuciscus cephalus (L.), using decapsulated Artemia as direct food

Little is known about the larviculture of the chub, Leuciscus cephalus (L.), an endangered cyprinid species endemic to European flowing waters. The use of decapsulated Artemia cysts as food for chub larviculture was investigated. After 3‐day feeding with the rotifer Brachionus calyciflorus, the larvae were fed on different diets: (i) dried decapsulated Artemia cysts, (ii) Artemia nauplii, (iii) rotifers for seven more days and then Daphnia collected from a pond, and (iv) an artificial diet. After a 24‐day rearing period, the highest survival rate was obtained with the larvae receiving decapsulated Artemia cysts. Feeding of the larvae with an artificial diet resulted in a significantly lower survival rate compared with the other groups. At the end of the experiment, the larvae fed on Artemia nauplii yielded a significantly higher mean length compared with the other groups. Feeding an artificial diet resulted in a significantly lower average weight and mean length gain compared with the other groups.