Genetic structure and phylogeography of European catfish (Silurus glanis) populations

The genetic structure of Silurus glanis (Europe's largest freshwater fish species) across most of its natural distribution was investigated using 10 microsatellite loci. The revealed levels of genetic diversity were much higher than previous allozyme and restriction fragment length polymorphism mitochondrial DNA analyses had shown; relative levels of variability among populations were however, in good agreement with the previous studies. Populations from large basins (Volga and Danube rivers) were the most polymorphic, while samples from the smaller Greek rivers, which are more prone to genetic bottleneck, exhibited the lowest levels of genetic diversity. Microsatellite multilocus genotyping permitted the assignment of individual fish to their population of origin with a score as high as 98.3%. Despite the great genetic differentiation of S. glanis populations, no consistent pattern of geographical structuring was revealed, in contrast to previous studies of European freshwater fish species. A model of isolation by distance seems more probable and a hypothesis of recent dispersion from only one glacial refugium is proposed. The discovery of the highest levels of microsatellite and mitochondrial diversity in the Volga sample and the presence of river connections, during the Pleistocene, between this area and all major areas of the present catfish distribution, place this refugium around the Ponto-Caspian region. Combining these data with those from previous studies, a number of markers are now available to monitor wild and hatchery populations even at the individual level.     

Chromosome banding study of European catfish,Silurus glanis (Pisces,Siluridae)

The karyotype of European catfish (Silurus glanis L.) was analyzed sequentially by means of silver staining and the chromomycin A₃ (CMA₃)/distamycin A (DA)/DAPI fluorescence technique and by C-banding, respectively. The nucleolus organizer regions (NORs) were localized on the submetacentric pair No. 14. Brilliant CMA₃ fluorescent heterochromatin blocks corresponded to the NORs visualized by silver staining. No DA/DAPI-bright positive fluorescent patterns were detected while C-banding led to the detection of specific banding patterns on several chromosome pairs.—Using these banding data, the karyotype of S. glanis was redescribed.     

Diel dualism in the energy consumption of the European catfish Silurus glanis

Twenty individuals of the largest European freshwater predator, the European catfish Silurus glanis, were tagged with electromyogram (EMG) physiological telemetry sensors. The fish were observed during diel cycles during the spring and summer in the Elbe and Berounka Rivers, Czech Republic. The purpose of this study was to determine whether diel dualism in the activity of S. glanis occurs naturally or is induced by the laboratory environment and by the conditions occurring in aquaculture. The results obtained from the riverine environment tended to show dualism in the use of the light and dark phases of the day because 35% of the individuals varied from a site‐specific common diel activity pattern. The EMG values increased in accordance with the mass (M) of the fish. To eliminate the influence of M on individual energy consumption, the EMG records were analysed in terms of the EMG:M ratios. High individual variability was found in these ratios. The diel activity pattern of the individuals with relatively high energy consumption differed from the common diel activity pattern. In contrast, the fish that adopted the common diel activity pattern displayed relatively low energy consumption. The results of this study indicated that dualism and energy consumption are related. The EMG values also varied with the values of the environmental variables. Increasing temperature was associated with high EMG values, whereas the EMG values decreased with increasing flow.     

Efficacy of tagging European catfish Silurus glanis (L., 1758) released into ponds

The efficacy and sub‐lethal consequences of single and double tagging European catfish Silurus glanis with Petersen disc and passive integrated transponder (PIT) tags were examined in short (laboratory) and longer‐term (field) experiments. Tag retention in the laboratory was 100%, with normal behaviour (i.e. feeding) in all fish returning within 36 h. In the field, 65 of 120 tagged S. glanis were recaptured from five small study ponds, with 85% retaining their PIT tags, though recapture rates and tagging efficacy were highly variable amongst locations. This is consistent with literature for other fishes, suggesting that tagging efficiency is variable across species and largely context dependent (fish length, tagging location, habitat).     

Application of DNA metabarcoding on faeces to identify European catfish Silurus glanis diet

In this study, the results of conventional stomach‐content analysis are compared with the recent DNA metabarcoding approach on faeces to identify fish species consumed by non‐native European catfish Silurus glanis in the Garonne River (south‐western France), with a special emphasis on anadromous prey. Fourteen prey species were identified in the stomach contents or faeces, including four anadromous fish species. Despite higher intestine than stomach emptiness, more species were identified through faecal analysis (11 of 14) than through stomach‐content analysis (five of 14) suggesting that DNA metabarcoding on faeces is an efficient, non‐intrusive technique to study the diet of predatory fishes.     

Mutual influence of protein and lipid feed content on European catfish (Silurus glanis) growth

We wished to determine protein and lipid content in pelleted raw fish food, necessary for optimal growth of European catfish (Silurus glanis). Experiments were set up in 20 cages, each holding 30 young catfish. Fishes in each cage received a different food combination over a 98‐day period at favourable physical and chemical water conditions. Food protein content varied between 37.5 and 45%, while lipid content, added in the form of soybean oil, varied between 3 and 11%. The oil contained an adequate ω‐fatty acids concentration, necessary for fish growth. The main growth indicators determined at the end of experiment were total body gain, specific growth rate and feed conversion ratio. There was a high statistical difference among the experimental groups receiving variable food combinations for each growth parameter (P < 0.001). Both protein and lipid food content affected growth parameters, but in a different manner. Further analysis – percentage of change depending on lipid to protein ratio and bivariate surface analysis – allowed us to recognize the most economical combination: 39.5% protein + 9% lipid content. The addition of 9% soybean oil to the fish food reduces the necessary protein concentration by 5.5%, with resulting identical catfish growth effects.     

Comparison of methods for hatchery-scale triploidization of European catfish (Silurus glanis L.)

Heat-, cold- and hydrostatic pressure shocks were applied in order to improve triploidy induction in European catfish ( Silurus glanis L . ). A 41°C heat shock (45 s, starting 9 min after gamete activation) provided 88% triploids and a high percentage of malformation (38.8 ± 4.1%). The superior 6°C cold shock (20 min, starting 9 min after gamete activation) gave 100% triploids and a 33.4 ± 3.8% triploid yield. The earliest hydrostatic treatments (600 kg cm²), lasting 4 min and starting 3 min after gamete activation, gave 97.8 ± 1.8% triploids and a 33.7 ± 16.9% triploid yield. The ploidy level was investigated using four approaches: karyotyping, quantification of Ag-stained nucleoli per cell, flow cytometry, and erythrocyte nuclear sizing by computer-assisted image analysis. Induction of triploidy under mass conditions in three experiments gave triploid percentages of 74%, 83% and 66%. Five months later, the percentage of triploids significantly decreased to 12.4%, 8.2% and 21.4%. The growth performance of yearlings was better in diploids than in triploids. Differences between diploids and triploids were 13.5% (NS), 27.6% (P   < 0.001) and 25.4% (P   < 0.05) in the three experiments. Analysis of variance showed the influence of ploidy (P   < 0.001) on growth rate, and multiple range analysis (LSD) assessed differences between total diploids (12.6 g) and total triploids (9.5 g) at the P   < 0.01 level.     

The influence of temperature on the growth of the European catfish (Silurus glanis L.)

The influence of temperature on growth, feed conversion, and protein and gross energy utilization was assessed in two experiments using the European catfish, Silurus glanis L. Animals of 55 g and 24 g were used in trials lasting 6 and 7 weeks, respectively. The optimum temperature for growth was within the range of 25 to 28°C, with the best results obtained at 27°C. Using à feed of low protein content (30.3%), a feed conversion of 1.67 was achieved at à specific growth rate of 1.78% of body weight/day.     

Electron microscopic demonstration of lectin binding sites in the taste buds of the European catfish Silurus glanis (Teleostei)

Taste buds in the European catfish Silurus glanis were examined with electron microscopic lectin histochemistry. For detection of carbohydrate residues in sensory cells and adjacent epithelial cells, gold-, ferritin- and biotin-labeled lectins were used. A post-embedding procedure carried out on tissue sections embedded in LR-White was applied to differentiate between the sensory cells: The lectins from Helix pomatia (HPA) and Triticum vulgare (WGA) bound to N-acetyl-galactosamine and to N-acetylglucosamine residues occurring especially in vesicles of dark sensory cells. This indicates a secretory function of these cells. Most light sensory cells--with some exceptions, probably immature cells--, are HPA-negative. The mucus of the receptor field and at the top of the adjacent epithelial cells was strongly HPA-positive. Pre-embedding studies were performed in order to obtain information about the reaction of the mucus with lectins under supravital conditions. The mucus of the taste bud receptor field exhibited intensive binding to WGA, but not to the other lectins tested. Most lectins bound predominantly to the surface mucus of the nonsensory epithelium and to the marginal cells close to the receptor field. The strong lectin binding to mucins and the relatively weak lectin binding to cell surface membranes in pre-embedding studies suggest that the mucus possibly serves as a barrier which is passed selectively only by a small amount of lectins or lectin-carbohydrate complexes. Lectin-carbohydrate interactions may play a role in recognition phenomena on the plasmalemmata of the taste bud sensory cells. Recognition processes directed to bacteria or viruses should be considered as well.     

Electron microscopic demonstration of lectin binding sites in the taste buds of the European catfish Silurus glanis (Teleostei)

Summary Taste buds in the European catfish Silurus glanis were examined with electron microscopic lectin histochemistry. For detection of carbohydrate residues in sensory cells and adjacent epithelial cells, gold-, ferritin-and biotin-labeled lectins were used. A post-embedding procedure carried out on tissue sections embedded in LR-White was applied to differentiate between the sensory cells: The lectins from Helix pomatia (HPA) and Triticum vulgare (WGA) bound to N- acetyl-galactosamine and to N-acetylglucosamine residues occurring especially in vesicles of dark sensory cells. This indicates a secretory function of these cells. Most light sensory cells — with some exceptions, probably immature cells —, are HPA-negative. The mucus of the receptor field and at the top of the adjacent epithelial cells was strongly HPA-positive. Pre-embedding studies were performed in order to obtain information about the reaction of the mucus with lectins under supravital conditions. The mucus of the taste bud receptor field exhibited intensive binding to WGA, but not to the other lectins tested. Most lectins bound predominantly to the surface mucus of the nonsensory epithelium and to the marginal cells close to the receptor field. The strong lectin binding to mucins and the relatively weak lectin binding to cell surface membranes in pre-embedding studies suggest that the mucus possibly serves as a barrier which is passed selectively only by a small amount of lectins or lectincarbohydrate complexes. Lectin-carbohydrate interactions may play a role in recognition phenomena on the plasmalemmata of the taste bud sensory cells. Recognition processes directed to bacteria or viruses should be considered as well.Parts of this investigation were presented at the XI. Annual Meeting of the Association for Chemoreception Sciences (AChemS XI), held at Sarasota, Fl, April 12–14, 1989 (Witt and Reutter 1989)     

Age and growth of the European catfish (Silurus glanis) in a Turkish Reservoir and comparison with introduced populations

The European catfish, Silurus glanis, is native to eastern Europe and western Asia and is among the largest freshwater fish in the world. Despite its increasing economic importance and its frequent introductions, the ecology and life-history of this species is poorly known due to the difficulty of sampling such a large species in large rivers and standing waters. Our study provides the first data on age and growth of this species in Turkish waters, where it is native. We report the length-weight relationships and age and size structure of this population, which were significantly different between females and males. A marginal increment analysis indicated that annulus formation occurred between May and June. The estimates of three growth functions (von Bertalanffy, logistic and Gompertz) are reported, with the von Bertalanffy growth providing a better fit and more realistic parameter estimates. Growth rates were significantly higher in males than in females and were overall higher compared to other native populations but similar to introduced populations of similar latitude.     

Purification and properties of serine proteinases from European catfish Silurus glanis L. pancreas

Three trypsin isoforms (designated as T1, T2, and T3), three chymotrypsin isoforms (Kh1, Kh2, and Kh3), and two elastase isoforms (E1 and E2) were isolated from the pancreas of European catfish Silurus glanis L. by salting out with (NH4)2SO4, gel chromatography on Sephadex G-75, and ion exchange chromatography on DEAE cellulose. Isoelectric points of the enzymes, determined by isoelectric focusing, amounted to 4.42 for T1, 5.64 for T2, 6.90 for T3, 4.93 for Khl, 5.23 for Kh2, 6.18 for Kh3, 6.17 for E1, and 8.48 for E2. Molecular weights of proteinases within each group were close and amounted to 30100 Da for trypsins, 39800 Da for chymotrypsins, and 24000 Da for elastases. The enzymes isolated displayed maximal activities at alkaline pH values. Inhibitor analysis demonstrated that all the proteinases isolated from European catfish pancreas belonged to the serine type.     

Susceptibility of European catfish (Silurus gianis) to Edwardsiella ictaluri

European catfish (Silurus glanis) were tested for their susceptibility to the bacterium Edwardsiella ictaluri. The LD₅₀ of E. ictaluri when injected into European catfish was 5.4 × 10⁶ compared to 7.1 times; 10⁴ for channel catfish (Ictalurus punctatus). E. ictaluri was isolated from dead and moribund European catfish and the bacterium was also detected in kidney smears by an indirect fluorescent antibody (FA) technique. The bacterium was not isolated or detected by FA from surviving fish 15 days after injection. No clinical signs of E. ictaluri infection were noted in European catfish, but these were prevalent in the channel catfish. These experiments indicate that under experimental conditions European catfish are not as susceptible to E. ictaluri as channel catfish.     

Linolenic acid supplementation in the diet of European catfish (Silurus glanis): effect on growth and fatty acid composition

In an 18‐week treatment, the effect of linolenic acid on European catfish (Silurus glanis L.) growth indicators was investigated as to weight gain, feeding coefficient (FCR value), specific growth rate (SGR value), protein efficiency ratio (PER value), productive protein value (PPV) and survival rate. Concurrently, the fishmeat chemical composition was also investigated. The experiment was organized into four groups, each divided into three subgroups. Stocked in each of 12 cages were 30 × 1‐year‐old catfish, with individual weights ranging from 148.5 to 151.5 g/ind. All fish were given standard feed for European catfish which contained 45% protein. The first batch, control group (C), received no additional linolenic acid. Linolenic acid was added to the feed of the second (E₁), third (E₂) and fourth group (E₃), at 0.5, 1.0 and 1.5%, respectively. Growth indicator improvement was best in the E₂ group fed the 1% linolenic acid, whereby the fish weight gain was 12.6% higher and the feeding coefficient 12.9% lower, while SGR, PER and PPV values were 6.1, 12.0 and 15.8% better than the control group. Growth indicators were also significantly (P < 0.01) improved in the three groups receiving additional linolenic acid in comparison to the control group. Moreover, this addition positively affected fishmeat quality by increasing meat protein content from 18.04% (C) to 18.79% (E₃). The total unsaturated fatty acid content also increased from 65.07% (C) to 69.82% (E₃), and the total saturated fatty acid content decreased from 31.36% (C) to 26.50% (E₃); consequently, the ratio of unsaturated to saturated fatty acids increased from 2.07% (C) to 2.63% (E₃). It can be concluded that the addition of 1% linolenic acid to standard catfish feed has beneficial effects on fish growth indicators and meat quality.     

Reduced genetic diversity and low effective size in peripheral northern European catfish Silurus glanis populations

Using 10 polymorphic microsatellites and 1251 individual samples (some dating back to the early 1980s), genetic structure and effective population size in all native and introduced Swedish populations of the European wels catfish or Silurus glanis were studied. Levels of genetic variability and phylogeographic relationships were compared with data from a previous study of populations in other parts of Europe. The genetically distinct Swedish populations displayed comparably low levels of genetic variability and according to one-sample estimates based on linkage disequilibrium and sib ship-reconstruction, current local effective population sizes were lower than minimum levels recommended for short-term genetic conservation. In line with a previous suggestion of postglacial colonisation from a single refugium, all Swedish populations were assembled on a common branch in a star-shaped dendrogram together with other European populations. Two distinct subpopulations were detected in upper and lower habitats of River Emån, indicating that even minor dispersal barriers may restrict gene flow for wels in running waters. Genetic assignment of specimens encountered in the brackish Baltic Sea and in lakes where the species does not occur naturally indicated presence of long-distance sea dispersal and confirmed unauthorised translocations, respectively.     

Susceptibility of European sheatfish Silurus glanis to a panel of ranaviruses

The current study was undertaken in order to assess the risk that different ranaviruses might impose on European sheatfish aquaculture. As the European sheatfish virus (ESV) is a known pathogen causing losses in European sheatfish aquaculture, it was assumed that closely related exotic ranaviruses might also be able to infect European sheatfish and probably cause disease and mortality in this species. The differential susceptibility of European sheatfish (Silurus glanis) to various ranavirus isolates was assessed at two different temperatures (15°C and 25°C) in a recirculation system. Fish were infected experimentally with a panel of ranavirus isolates including ESV, European catfish virus (ECV), European catfish virus isolate 24 (ECV‐24), Epizootic haematopoietic necrosis virus (EHNV), Rana esculenta virus isolate Italy 282/ I02 (REV), short‐finned eel virus (SERV), Bohle iridovirus (BIV), guppy virus 6 (GV6), doctor fish virus (DFV) and Frog virus 3 (FV3). Significant mortalities were observed, as expected, in fish infected with ESV at 15°C (100%) as well as at 25°C (86/83%). Fish infected with ECV at 15°C showed no clinical signs of disease (8% mortality), whereas those fish infected at 25°C exhibited a cumulative mortality of 54%. Fatal disease was also induced by Italian isolate ECV‐24 at 25°C (81%). Virus isolates ESV, ECV and ECV‐24, generally the most genetically closely related viruses, were successfully isolated from dead fish by cell culture with subsequent identification by polymerase chain reaction (PCR) and sequence analysis. However, no mortality or clinical signs of disease were observed in the groups of sheatfish infected with the other ranaviruses investigated in the study, and none of those viruses were re‐isolated in cell culture or identified by PCR. It was concluded that European sheatfish are susceptible to infection with ESV, ECV and ECV‐24 under laboratory conditions, but not to infection with EHNV, REV, SERV, BIV, GV6, DFV or FV3. For ESV, the incubation period was shorter at 25°C compared to 15°C water temperature, but whereas all fish died after ESV infection at 15°C, some fish survived the infection at 25°C. Futhermore, the very young sheatfish were susceptible to ECV and ECV‐24 at 25°C, whereas there was no significant mortality in the group of older sheatfish challenged with ECV at 15°C. Therefore, the clinical characteristics of the disease seem to depend on the age of the fish as well as on the water temperature.     

Mitochondrial DNA variation in European populations of Silurus glanis

Mitochondrial DNA diversity of 13 wild Silurus glanis populations (covering the entire range of the species) and eight hatchery populations was investigated. PCR-RFLP analysis of four regions of mitochondrial DNA (cytochrome b, control region, ND-5/6) was used. Nineteen haplotypes were found. Thirteen of them were private. The proportion of total genetic diversity attributable to population differentiation was almost 80%. Despite the existence of significant differentiation between populations for mtDNA variation, no consistent pattern of geographic structuring was revealed and nucleotide divergence among S. glanis populations was low. These phenomena are discussed with regard to the impact of glaciation events. The domesticated stocks show less genetic diversity than natural ones, possibly due to their mode of management. Analysis of three European catfish species S. glanis, S. aristotelis and Silurus triostegus (sampled in the Euphrates river) revealed several endonucleases which produced restriction phenotypes diagnostic for the three species.     

Explicit site fidelity of European catfish (Silurus glanis,L., 1758) to man‐made habitat in the River Meuse,Netherlands

After a century of near absence, records of European catfish (Silurus glanis, L., 1758) in the Netherlands have increased markedly since 1994. The majority of records (about 60% of the total) originate from a 12 km dammed section of the River Meuse in the province of Limburg. This paper reports on a study conducted to obtain information on the behaviour and habitat preferences of catfish at this location, whereby 20 adult Silurus glanis were implanted with both acoustic and transponder tags and their behavior monitored using a combination of passive and active methodologies. All fish remained within the study area and did not enter either the contiguous riverine sections or connected lakes. The catfish expressed schooling, nocturnal activity and strong site fidelity to a specific river section containing artificial habitat (in the form of moored boats) and were influenced by the presence and operation of nearby energy generation plants. The results are discussed in relation to the potential growth and dispersal of catfish populations and the extent to which anthropogenic activities (particularly energy generation schemes) at the site have inadvertently created favourable habitat that has assisted the colonization of catfish and impacted negatively on the wider fish community.     

Cryopreservation of European catfish Silurus glanis sperm: sperm motility, viability, and hatching success of embryos

The aim of this study was to elaborate cryopreservation methods for ex situ conservation of European catfish. The success of sperm cryopreservation was evaluated by post-thaw sperm motility and velocity, percentage of live spermatozoa and fertility (hatching rates) using frozen/thawed sperm. The best hatching rates of 82-86% were obtained with sperm stored for 5 h before freezing in immobilizing solution and frozen with Me2SO in concentrations of 8, 10, and 12%, or with a mixture of 5% Me2SO and 5% propandiole. These results did not significantly differ from the fresh sperm control sample. The percentage of live spermatozoa in frozen/thawed sperm did not correlate with hatching rate or motility of spermatozoa, but was negatively correlated with velocity of spermatozoa (r=-0.47, P=0.05). The percentage motility in frozen/thawed sperm ranged from 8 to 62%, when sperm was stored in immobilizing solution 5h before freezing. The average value in the fresh sperm (control) was 96%. The frozen/thawed sperm motility rate significantly correlated with the hatching rate (r=0.76, P=0.0002), but not with the percentage of live spermatozoa (r=0.16, P=0.52) or the sperm velocity (r=0.07, P=0.79). The velocity of frozen/thawed spermatozoa ranged from 37 to 85 microm/s, whereby methanol concentrations of 7.5 and 10% resulted in highest velocities. Freezing sperm volumes of 1-4 ml did not affect the quality of frozen/thawed sperm.