Additive, dominance genetic effects for growth-related traits in common carp, Cyprinus carpio L.

Additive, dominance genetic effects were analysed for body weight (BW) and seven morphometric traits from three variants of red common carp, Cyprinus carpio var. singuonensis, C. carpio var. wuyuanensis and C. carpio var. color, based on a diallel cross‐mating design by using the additive–dominance genetic model. The results indicated that at 8 months of age (juvenile stage), the traits of pre‐dorsal height (PDH), pre‐dorsal width (PDW) and caudal peduncle height (CPH) were mainly controlled by additive effects; the traits of BW, total length (TL), standard length (SL), head length (HL) and caudal peduncle length (CPL) were mainly controlled by dominance effects. The narrow‐sense heritabilities of all traits ranged from 0.000 to 0.556; however, the broad‐sense heritabilities ranged from 0.453 to 0.775. At 20 months of age (adult stage), the traits of TL, SL, PDH and CPL were mainly controlled by additive effects, whereas the traits of BW, PDW, HL and CPH were mainly controlled by dominance effects. The narrow‐sense heritabilities of all traits ranged from 0.000 to 0.710; the broad‐sense heritabilities ranged from 0.629 to 0.934. Through the genetic merit prediction from three parents, it was found that the additive and dominance effects of C. carpio var. color could increase the BW and body length in their progeny, but these two genetic effects of C. carpio var. singuonensis and C. carpio var. wuyuanensis could decrease BW in their progeny. The genetic effects of C. carpio var. wuyuanensis could reduce total length, SL and caudal peduncle length, but increase body height in their progeny.     

Temperature-dependent activity of brackish water yellow eels, Anguilla anguilla L.

Abstract. Female yellow eels, Anguilla anguilla L., were caught with fyke nets in the brackish water bight Hallangspollen, Norway during 1983 and 1984. Fishing was performed continuously for a variable number of days (defined as the fishing period), and the nets were monitored at uneven intervals during the fishing period. The eel activity, expressed as catch per unit effort (CPUE), was positively correlated (R²= 0·74; P < 0·001) with water temperature (defined as a regression of temperature variation during the fishing period). The multiple regression model of CPUE on water temperature was improved by including the number of fishing days in each fishing period in the regression model (R²= 0·91; P < 0·001). With high fishing efforts the predicted CPUE decreased below that which was predicted by the temperature model alone.     

Vaccine development for winter ulcer disease, Vibrio viscosus, in Atlantic salmon, Salmo salar L.

Coldwater Vibrio species isolated from Atlantic salmon, Salmo salar L., during winter ulcer disease outbreaks at saltwater sites in Norway and Iceland were characterized phenotypically, tested for virulence, and used to evaluate the efficacy of multivalent, oil-adjuvanted vaccines. The intraperitoneal (i.p.) injection of rainbow trout, Oncorhynchus mykiss (Walbaum), in fresh water with one bacteria species isolated during winter ulcer outbreaks, V. ‘viscosus’, produced rapid mortality and disease signs which resembled those observed during natural outbreaks [10⁵ colony-forming units (cfu) fish^??1]. Another species, V. ‘wodanis’, was not virulent to rainbow trout (10³–10⁶ cfu fish^??1). Although vaccination of rainbow trout with a mineral-oil-adjuvanted, injectable vaccine containing V. anguillarum (serotypes 01 and 02), V. salmonicida and Aeromonas salmonicida did not provide protection against injection challenge with V. viscosus, vaccines which included V. viscosus produced significant protection in Atlantic salmon and rainbow trout. Atlantic salmon vaccinated with an oil-adjuvanted vaccine containing V. viscosus, V. wodanis and atypical A. salmonicida produced a relative percentage survival (RPS) of 97% when challenged i.p. with V. viscosus, demonstrating cross-protection between strains from Iceland and Norway. Short-term efficacy was demonstrated in rainbow trout by injection challenge at 21 and 43 days post-vaccination with an oil-adjuvanted vaccine containing V. viscosus, V. anguillarum (01/02), V. salmonicida and A. salmonicida, which produced an RPS of 96–99%. Rainbow trout challenged with V. viscosus at 52 and 362 days post-vaccination produced an RPS of 93% and 79%, indicating that vaccination provided long-term protection. In a similar manner, rainbow trout injected i.p. with 0.2 mL of a vaccine containing the five bacteria species and infectious pancreatic necrosis virus produced a 90% RPS when challenged with V. viscosus 66 days later. The high RPS under a severe challenge burden, along with disease signs in experimental freshwater challenges which resembled the saltwater disease condition, indicated that V. viscosus is a contributing factor to winter ulcer and that vaccination will protect against the disease.     

Estimation of gut contents of freshwater mussel, Lamellidens marginalis L.

This study examined the food organisms found in the gut of freshwater mussels, Lamellidens marginalis L. A total of 34 taxa of food organisms were recorded, out of which 30 taxa belonged to phytoplankton and four taxa to zooplankton. Both the groups comprised three families each: Cyanophyceae (blue‐green algae), Chlorophycea (green algae) and Bacillariophyceae (diatoms) constituted phytoplankton, whereas Euglenaceae, Chlamydodontidae and Brachionidae belonged to zooplankton. The total number of frequencies of phytoplankton (n=537) were almost nine times more than that of zooplankton (n=60). When blue‐green algae, green algae, diatoms and zooplankton (all the three families together) were tested for significant difference (P<0.05) following Duncan's multiple range test, the result showed only two groups. Blue‐green algae, green algae and zooplankton were not significantly different (P<0.05), forming group A, whereas diatoms were significantly different from others, forming group B. The present study showed that the maximum number of organisms that mussels feed upon belong to green algae, followed by diatoms, blue‐green algae and few taxa of zooplankton.     

Bacterial septicaemia in prerecruit edible crabs,Cancer pagurus L.

Juvenile edible crabs, Cancer pagurus L., were surveyed from Mumbles Head and Oxwich Bay in South Wales, UK, and the number of heterotrophic bacteria and vibrios in the hemolymph was determined. The percentage of crabs with hemolymph containing bacteria was variable over the survey with higher numbers of animals affected in summer than in winter. Post‐moult crabs contained significantly higher numbers of heterotrophic bacteria in the hemolymph than pre‐ and intermoult animals. Crabs with cuticular damage to the gills also had significantly higher numbers of bacteria in the hemolymph. Crabs were found to have a high prevalence of infection by the dinoflagellate, Hematodinium. Such animals had significantly fewer bacteria in the blood in comparison with Hematodinium‐free animals. Of the 463 crabs surveyed, only 3 individuals had hemolymph containing 2000 + CFU mL^?1. Based on 16S rRNA gene sequences, two of these crabs contained a Vibrio pectenicida‐like isolate, while the other had a mixed assemblage of vibrios. Although 59% of the crabs surveyed had culturable bacteria in the hemolymph, the majority only had small numbers (<2000 CFU mL^?1), suggesting that such infections may be of limited importance to the sustainability of the crab fishery in this region.     

Assessment of flesh colour in Atlantic salmon, Salmo salar L.

The degree of pigmentation in muscle of Atlantic salmon, Salmo salar L., fillets of fish that were fed eight diets fortified with 10, 20, 40, 60, 80.100, 150 and 200 mg astaxanthin kg^?1 and a non-supplemented control diet from 3 to 21 months was assessed using different methods. A tristimulus colorimeter (Minolta Chroma Meter) was used to measure the colour composition of the fillets instrumentally. The colour was also determined using the Roche Colour Card for Salmonids. The concentration of astaxanthin in the muscle was measured by chemical analyses. All measurements for colour were done directly on the epaxial muscle anterior to the dorsal fin. The lightness factor (L *). the red/green chromaticity (a*), the yellow/blue chromaticity (b*) and the saturation C* of the colorimetric readings and the Colour Card scores were compared with the chemical analyses. The astaxanthin concentration in the flesh varied from 1 to 10 mg kg^?1 and the visual appearance of the fillets varied from yellowish-white to red. The relationship between the a*, b* and C* values and the astaxanthin concentration in the muscle was non-linear. Non-linear regression lines were found between the a* value and the astaxanthin concentration in the flesh (r²= 0.974) and the b* value and the astaxanthin concentration in the flesh (r²= 0.984). The instrument was not able to detect differences in astaxanthin concentration at astaxanthin levels above 3-4 mg kg^?1 using the presented method directly on the fillet. The instrument might be useful for rejecting groups of salmon with poor pigmentation. A good linear regression was found between the Colour Card score and the mean astaxanthin concentration in the flesh (r² - 0.992). The Colour Card provided a better prediction of the astaxanthin concentration at higher astaxanthin levels than the Chroma Meter. None of the methods provided a satisfactory prediction of the astaxanthin concentration in the muscle of individual fish using the presented methods.     

Modelling growth in cultures of Oreochromis niloticus (L.) and Cyprinus carpio L. in Pernambuco, Brazil

Different growth models in cultures of Oreochromis niloticus (L.) and Cyprinus carpio L. were compared in Pernambuco, Brazil. The Chapman–Richards growth model was shown to be the most appropriate for Oreochromis niloticus, while Silva's model best expressed growth for Cyprinus carpio.     

Use of duckweed, Spirodela polyrrhiza L. Schleiden, as a protein feedstuff in practical diets for tilapia, Oreochromis niloticus L.

The use of solar-dried duckweed, Spirodela polyrrhiza L. Schleiden, as a dietary protein component for tilapia, Oreochromis niloticus L., reared in glass tanks was evaluated. Six isonitrogenous diets (30% crude protein) were fed to all-male tilapia fingerlings for 56 days. The fish meal protein in the diets was substituted at a rate of 5%, 10%, 20%, 30% and 100% with duckweed. A diet without the duckweed served as a control. Growth performance and nutrient utilization of fish were based on daily weight gain, specific growth rate, feed conversion ratio, protein efficiency ratio and protein productive value. There were no significant differences (P > 0.05) in growth performance and nutrient utilization of fish fed on diets containing up to 20% duckweed inclusion and the control. However, increases in dietary duckweed inclusion resulted in progressively reduced growth performance and nutrient utilization of fish. Diet without fish meal (100% duckweed) gave the poorest result. The most cost-effective diet in terms of cost per unit gain in weight of fish was obtained with 30% duckweed dietary inclusion. The result showed that solar-dried up to 30% duckweed dietary inclusion as a replacement for fishmeal in practical diets supported fish growth and was cost-effective.     

Mitochondrial DNA variation in two samples of northern pike, Esox lucius L.

The mitochondrial genotype of northern pike, Esox lucius L., was characterized by restriction analysis performed over the entire mtDNA molecule (33 fish) and over a ～1.4-kb PCR-amplified segment of a mitochondrial non-coding control region (six fish). Out of eight restriction endonucleases used in the present study of the entire mitochondrial genome, the Nco I and Pst I enzymes produced variant fragment patterns which allowed identification of three mitochondrial genotypes. Low mtDNA diversity was shown by estimates of pairwise nucleotide substitution values (d; mean ± SE = 0.49 ± 0.43%), haplotype diversity (h= 0.16) and the nucleotide diversity index (π= 0.06%). The present authors developed a PCR protocol for mitochondrial DNA non-coding control region to search for more variation. Whereas the PCR assays revealed distinct length variation in this mtDNA segment, restriction analysis performed with three additional restriction enzymes showed no variability which could be attributed to a gain or loss of a particular restriction site. Although the mtDNA analysis proved ineffective in genetic discrimination between the two northern pike samples studied, fish with rare mtDNA genotypes could be used for conducting experimental genome engineering or stocking studies with this species.     

Cryopreservation of sperm of common carp, Cyprinus carpio L.

Milt of common carp, Cyprinus carpio L. was cryopreserved in pellet form with the use of 12 extenders. Most efficient were: BE2 original extender (containing 85 mM NaCl, 50 mM KCl, 3 mm CaCl₂, 1 mm MgCl₂ with 10% dimethyl-acetamide (DMA) and 10% addition of hen's egg yolk) and Kurokura et al.'s extender with 15% DMA and 10% yolk (about 73% and 69% of eyed eggs, about 61% and 52% of swim-up larvae, respectively). Within the most effective treatments, survival from the eyed-egg stage to the swim-up stage was similar to that observed in the control group. Survival from the eyed-egg stage to the swim-up stage (percentage of eyed eggs was considered as 100%) was highly significantly and positively correlated with the actual rate of swim-up larvae.     

Effect of feeding Spirulina platensis on the growth, proximate composition and organoleptic quality of common carp, Cyprinus carpio L.

The effect of feeding Spirulina platensis on the growth, carcass composition, organoleptic quality, digestive enzyme activity and digestibility of common carp, Cyprinus carpio L., was studied through a culture trial lasting 120 days. Four experimental diets were employed by replacing fish meal protein from the standard diet at 25%, 50%, 75% and 100% through the incorporation of Spirulina. Another diet with Spirulina as the sole source of protein was also used. The final weight gain, specific growth rate, food conversion ratio and protein efficiency ratio of common carp were not affected by Spirulina supplementation. However, the diet with Spirulina as the sole source of protein resulted in better net protein retention. The muscle RNA:DNA ratio of fish fed Spirulina diets was higher than that of fish fed control diet. There was no significant difference in carcass moisture and protein content in the fish fed Spirulina diets as compared to fish-meal-based control diet. The carcass ash and fat contents were positively and negatively correlated with dietary Spirulina level, respectively. Organoleptic evaluation revealed no effect of Spirulina feeding on the quality of both raw and cooked fish. The gut digestive enzyme activity did not show any definite trend with respect to Spirulina supplementation. Spirulina improved the protein digestibility of the diets.     

River bed construction: impact and habitat restoration for juvenile Atlantic salmon, Salmo salar L., and brown trout, Salmo trutta L.

The River Soya, Norway, was canalized for agricultural purposes. In order to compensate for damage to the Atlantic salmon, salmo salar L. and brown trout, Salmo trutta L. populations, different weirs were built. The aims of this study were firstly to analyse the effects of weirs covering the river bank and entire river bottom with blasted stones and secondly, to analyse the effects of sediments transported by freshets on the downstream salmon and trout populations after canalization. Restoration of the river bottom with blasted stones provided salmon with more substrate spaces. Densities of trout increased after the river bank was covered with stones. Sediments transported downstream from the canalized river stretch decreased the densities of juvenile salmon and trout.     

The First Results on Relationships Among Amphimictic Diploid, Diploid Gynogenic and Triploid Tench, Tinca tinca L. under Communal Testing

Amphimictic diploid (2n), induced meiotic gynogenic (MeiG) and induced triploid (3n) siblings of tench, Tinca tinca L., were separately nursed, marked and kept in long-term communal testing in pond until sex maturation and natural spawning. Survival rates during the initial periods were lesser in 2n and 3n compared to MeiG population (65%). The first overwintering period showed decreased survival of 3n (62.67%) compared to MeiG (90.0%) and 2n populations (91.67%). Survival rates during the next seasons ranged in 70.50–90.97% similarly in both 3n and 2n while in MeiG population it dropped to 38%. Weight and length growth of all populations studied showed significantly higher growth of 3n females and of 3n in general, compared to 2n and MeiG populations (P < 0.001). Health examinations found metacercaria of Diplosthomus spathaceum in eyes of 2-year-old 3n specimens, weak polyparasitic infections of skin and gills by Dactylogyrus, Trichodina, Thrichophrya, Apiosoma and Chilodonella spp. in 3n and 2n specimens but medium strong to strong infections of 100% of MeiG fish which might contribute to their enhanced mortality. Ploidy analysis of progeny obtained from spontaneous spawning of the broodstock showed 100% diploidy, indicating the following possibilities: 2n population might spawn itself, 2n males might also mate to gynogens, or 3n males might also mate to 2n and/or MeiG females causing spontaneous gynogenesis. Further pedigree analyses will be focused to highlight these questions.     

Folate in eggs and developing larvae of Atlantic halibut, Hippoglossus hippoglossus L.

Folate mobilization from the yolk compartment during larval development was studied by analysing the folate concentration in whole body, embryo and yolk in a single batch of Atlantic halibut, Hippoglossus hippoglossus L., eggs and larvae that showed successful fertilization and development. There was a net loss of approx. 50% of folate from yolk during endogenous feeding. Further, only 23% of the decrease in yolk folate was retained in the larval body. The data suggest a need for folate for metabolic and growth purposes during embryogenesis of approximately 2 μg g^?1 weight gain. Relative to these data and published folate requirement for cold‐water species, batches of egg from 16 Atlantic halibut brood fish contained variable and, for some batches, critically low levels of folate. This may constitute a potential problem for larval development until start feeding.     

Gillnet selectivity to pikeperch, Stizostedion lucioperca (L.), and perch, Percafluviatilis L., caught mainly wedged

Abstract. The size selectivity of gillnets to pikeperch, Stizosiedion lucioperca (L.), and perch, Perca fluviatilis L., caught mainly wedged, was estimated by the selective use of the deterministic model of Holt (1963). The selectivity characteristics for pikeperch based on samples in gillnets with 48-65-mm bar mesh were: k= 0·903 and σ= 4·93cm (fork length). When a higher efficiency of large-meshed nets is accounted for, the estimated selectivity curve shifts to the right. The condition of the pikeperch had a major influence on the position and shape o(the selectivity curve. The selectivity to perch was established with 25–35-mm gillnets in a year with a poor food supply: k= 0·723 and σ= 2·59cm (fork length). Estimates based on literature data from an exploited perch population showed a much lower k= 0σ639 (total length) (σ= 2·69cm).     

Recovery of Hafnia alvei from diseased brown trout,Salmo trutta L., and healthy noble crayfish,Astacus astacus (L.), in Bulgaria

Hafnia alvei was isolated in Bulgaria from healthy noble crayfish, Astacus astacus (L.), and then from farmed diseased brown trout, Salmo trutta L., with signs of haemorrhagic septicaemia. The isolates were identified initially with conventional phenotyping and commercial Merlin Micronaut and API 20E rapid identification systems, followed by sequencing of the 16S rRNA gene. Hafnia alvei Bt1, Bt2 and Aa4 were of low virulence to rainbow trout and brown trout, although cytotoxicity was demonstrated by Bt1 and Bt2, but not by Aa4.     

Dietary selenium requirement for juvenile cobia,Rachycentron canadum L.

A 10‐week feeding trial was conducted to estimate the optimum dietary selenium (Se) requirement for juvenile cobia, Rachycentron canadum L. The basal diet was formulated to contain 50.6% crude protein from vitamin‐free casein, gelatin. A control diet (no added seleno‐dl ‐methionine) and five experimental diets containing 0.20, 0.40, 0.60, 0.80 and 1.00 mg seleno‐dl ‐methionine kg^?1 were prepared. Each diet was randomly fed to triplicate groups of juvenile cobia with initial weight 6.27±0.03 g in a flow‐through system. The Se concentration in rearing water was monitored during the feeding period, and was not detectable. The dietary Se level significantly influenced the survival, specific growth rate (SGR), feed efficiency and the Se concentrations in the whole body and vertebra of cobia. The Se‐dependent glutathione peroxidase (EC 1.11.119) activity increased with an increase in the dietary Se levels (P<0.05). Hepatic glutathione reductase (EC 1.6.4.2) activity was the highest in fish fed the diet with 0.21 mg Se kg^?1, and declined with an increase in the dietary Se levels. Based on broke‐line regression of SGR, the Se concentration in the whole body and vertebra, the Se requirements of juvenile cobia were 0.788, 0.811 and 0.793 mg Se kg^?1 diet in the form of seleno‐dl ‐methionine respectively.     

Induced spermiation in 3-year-old sterlet, Acipenser ruthenus L.

Spermiation in 3‐year‐old sterlet, Acipenser ruthenus (L.), males maintained under warm water conditions was induced by intramuscular injection of either (i) (D‐Ala⁶)‐GnRH‐ProNHEt (Kobarelin); (ii) mammalian GnRH analogue+metoclopramide (Ovopel); or (iii) human chorionic gonadotropin (Biogonadyl). The volume of milt, sperm concentration and motility were measured. A higher percentage of spermiating males was obtained after Kobarelin or Ovopel treatment (81.8% and 77.7% respectively) in comparison with fish treated with Biogonadyl (40.0%). However, only stimulation with Ovopel guaranteed motile spermatozoa in all spermiating males. Moreover, treatment with Ovopel resulted in the highest average milt volume, sperm concentration and motility. The average total number of motile spermatozoa (milt volume×sperm concentration×sperm motility) per individual was 0.99×10⁹, 5.31×10⁹ and 0.02×10⁹ after stimulation with Kobarelin, Ovopel or Biogonadyl respectively. Our data indicate that Ovopel is a good stimulator of spermation in sturgeons. Moreover, the time of sexual maturation could be significantly reduced in sturgeons maintained in cages under warm water conditions.     

In vitro passages impact on virulence of Saprolegnia parasitica to Atlantic salmon,Salmo salar L. parr

The effect of serial in vitro subculturing on three pathogenic strains of Saprolegnia parasitica was investigated. The isolates were passed through Atlantic salmon, Salmo salar L. parr, and then re‐isolated as single spore colonies. All strains caused infection. The isolate obtained from diseased fish served as a virulent reference culture and was designated ‘AP’ (‘activated through passage’). Successive subculturing was made by obtaining an inoculum from AP to produce the 2nd subculture and then passaged to the 3rd subculture (from the 2nd), until the 15th passage was obtained. Spores used to produce storage cultures were collected at passages 5, 10 and 15. The different passages of each strain were used to artificially infect Atlantic salmon parr. Morphological characterization of growth patterns was performed to observe differences occurring due to serial in vitro subculturing. Two of the strains declined in virulence after 15 successive in vitro subcultures, whereas one did not. This study is the first to investigate attenuation of virulence in Saprolegnia and whether or not isolates of S. parasitica should be passed through the fish host prior to challenge experiments. It reveals that some strains degenerate more rapidly than others when subjected to successive in vitro subculturing on glucose–yeast extract.     

Atypical Aeromonas salmonicida infection in naturally and experimentally infected cod, Gadus morhua L.

Cod, Gadus morhua L., of wild origin, were reared at different temperatures for 12 months. During this period, moribund and newly dead fish were examined and samples collected for bacteriology and histopathology. Atypical Aeromonas salmonicida was isolated from 10 individuals reared at or above 7 °C. The isolates were homogeneous with respect to biochemical and antibiogram characters and similar to the ssp. achromogenes National Collection of Industrial and Marine Bacteria, UK, type strain 1110 and reference strains that have been isolated from salmonids and haddock in Iceland. Histopathological analysis of the naturally infected cod showed typical ulceration associated with atypical A. salmonicida infection and also widespread granulomatous formations. One‐year‐old cod of farmed origin, kept at 9 °C, received intraperitoneal or intramuscular injection with different doses of atypical A. salmonicida, isolated from the above wild cod. Mortalities were monitored for 28 days and the LD₅₀ calculated. The route of bacterial injection influenced the mortality rate and LD₅₀ value and affected, to some extent, the pathological changes observed and humoral immune parameters. Pathological changes, including haemorrhage, early stages of granuloma formation and necrotic changes, were seen in several organs. Infection appeared to induce non‐specific antibody activity against trinitrophenyl (TNP)‐haptenated protein and may have activated the complement system. Specific antibody response against atypical A. salmonicida was not detected.