The purification and partial characterization of carp,Cyprinus carpio,vitellogenin

A procedure is described for the isolation of intact vitellogenin (c-VTG) from the carp, Cyprinus carpio. VTG was induced in juvenile females using oestradiol-17β and purified from the plasma using a combination of gel-filtration chromatography on Sepharose 6B and ion exchange chromatography on DEAE-cellulose. Purification procedures were conducted at low temperatures (below 9°C) in the presence of the proteolytic enzyme inhibitor aprotinin to prevent degradation. Intact c-VTG had an apparent molecular mass of 390,000 Daltons, but when extracted from plasma in the absence of aprotinin it underwent proteolysis into at least 2 protein fragments (apparent molecular masses of 230,000 and 96,000 Daltons), showing an instability of the native dimer. An amino acid analysis of c-VTG showed that its composition was almost identical to goldfish VTG, a species closely allied to the true carps and also similar to other oviparous vertebrate VTGs. Collectively, these data indicate that using these purification procedures VTG from carp, and probably other teleost species, can be isolated in an intact, highly purified form.     

Age and growth of yellow eel, Anguilla anguilla (L.), determined by two different methods

Abstract– Ageing and calculation of growth rate of eels has always been controversial, and remarkably little validation has been reported. This article describes the growth of yellow eels from three lakes in the Burrishoole system, Ireland, as determined by back-calculation from otoliths prepared by burning and cracking. Eels captured by fyke net ranged in length from 25.0 cm to 98.5 cm and the ages ranged from 6 to 49 years. Back-calculated growth was slow, with mean annual increments of 2.08 cm, 1.40 cm and 1.97 cm for the three lakes. Yellow eels captured in the Burrishoole system in 1987 and 1988 were marked by Pan Jet inoculated alcian blue dye and released back into the three lakes. Recaptures made over the following three years yielded considerable growth data. Mean annual growth rates recorded were 2.30 cm, 1.07 cm and 0.97 cm. These data compare favorably with the calculated growth rates, thereby validating the otolith analysis for the eels in the Burrishoole system.     

Biochemical and pharmacological characterization of adenosine A1 receptors in eel (Anguilla anguilla) brain

N⁶-cyclohexyl[³H]adenosine ([³H]CHA) was used to label adenosine A1 receptors in membranes prepared from male and female eel whole brain. The A1 receptor agonist [³H]CHA bound saturably, reversibly and with high affinity (K_d = 0.91 ± 0.12 nM; B_max = 120.36 ± 5.2 fmol mg⁻¹ protein). In equilibrium competition experiments, the adenosine agonists and antagonists all displaced [³H]CHA from high-affinity binding sites with the rank order of potency in displacing, characteristics of an A1 adenosine receptor. Mg²⁺ dramatically increased the affinity of [³H]CHA without modifying the maximal binding capacity. The specific binding was inhibited by guanosine 5′-triphosphate (K_i = 2.54 ± 0.98 μM). The [³H]CHA binding sites are ubiquitously distributed with a maximum in cerebellum and a minimum in olfactory bulb. No difference was observed between male and female brain. In eel brain, synaptosomes (P2), stimulation of adenosine 3′,5′-monophosphate (cyclic AMP) accumulation with 10⁻⁵ M forskolin was markedly reduced (45.5%) by treatment with the adenosine A1 receptor agonist CHA (10⁻⁴ M), and the reduction was reversed in presence of the selective A1 receptor antagonist 8-cyclopentyltheophylline (10⁻⁵ M). In superfused eel cerebellar synaptosomes, K⁺ stimulated the release of adenosine in a partially Ca²⁺-dependent manner. The findings, taken together, suggest the hypothesis that adenosine A1 receptors present in eel brain could modulate synaptic transmission, as A1 receptors do in other vertebrates.     

Enhancement of non‐specific immune responses in common carp,Cyprinus carpio,by dietary carotenoids obtained from shrimp exoskeleton

The immunostimulatory role of carotenoid extract from shrimp processing discards was evaluated by feeding common carp fingerlings with a diet containing carotenoid extract for 21 weeks at 100 and 200 mg kg^?1 astaxanthin levels. Haemoglobin content was significantly (P < 0.05) higher in the blood of fish fed with carotenoid diet (>8.1 g dL^?1) compared with that from fish fed with carotenoid deficient diet (6.86 g dL^?1) and also leukocyte counts were higher (P < 0.05). No differences (P > 0.05) were observed in total serum protein, globulin level and albumin‐globulin ratio, but albumin content was higher (P < 0.05). Respiratory burst activity was significantly (P < 0.05) higher, the serum lysozyme activity almost doubled and the serum bactericidal activity was significantly increased when the fish were fed with diet containing 200 mg kg^?1 of astaxanthin but no significant differences were observed in serum trypsin inhibitory activity. There was a significant (P < 0.05) increase in leukocyte myeloperoxidase activity due to dietary carotenoids. Challenging fish with Aeromonas hydrophila after the feeding period resulted in 50% mortality in the control group while in the group fed with diet containing 100 mg kg^?1 astaxanthin, the mortality rate was 15%. No mortality and even symptoms of infection was not observed in the group fed with diet containing 200 mg kg^?1 of astaxanthin. The study indicated that carotenoid extract from shrimp processing discards can effectively be used as immunostimulants in aquaculture of carps and dietary carotenoids were found to enhance various immune defence mechanisms and also provide protection against the infection of pathogen A. hydrophila.     

Estuarine and fluvial recruitment of the European glass eel, Anguilla anguilla, in an exploited Atlantic estuary

A glass eel fishery exists downstream of the Arzal estuarine dam in the Vilaine (Brittany, France). Catch statistics were collected between 1996 and 2000, and processed using a subsampling technique which allowed data from a reliable subsample to be extrapolated to the whole fishery. During the same period, glass eel migration into fresh water was monitored using a glass eel trap located on the dam – the upstream limit of the fishery. The analysis of glass eel biology and exploitation shows that the glass eel fishery is very intensive and that there was more or less no escapement during the fishing seasons studied. The proportion of the stock successfully migrating towards fresh water, as compared with the total catch is estimated to range between 0.3 and 3.9%. In such a fishery, the fishing effort affects the abundance. Consequently, the total catch has to be used instead of catch‐per‐unit‐effort to estimate abundance.     

Differences in head shape of the European eel, Anguilla anguilla (L.)

Differences in eel, Anguilla anguilla (L.), head shape were the result of greater increases in head width, rather than changes in head length. The ratio of head width:total length (HW:TL) increased significantly ( t -test, P < 0.05) from the glass-yellow eel stage. Cultured yellow eels were exclusively narrow headed (mean HW:TL = 0.027, range = 0.023–0.032), while wild yellow eels displayed an array of head shapes (mean HW:TL = 0.033, range = 0.023–0.046). Therefore, broad heads (HW:TL 0.033) occurred only among wild yellow eels sampled and may have resulted from diet. Cultured yellow eels consumed only small pellet material. Of wild yellow eels stomachs containing food, 78% of broad-headed eels consumed large and/or hard-bodied organisms (e.g. beetles, fish, molluscs and Notonecta sp.), while 83% of narrow-headed eels consumed exclusively small/soft-bodied prey (e.g. amphipods and chironomids).     

cDNA cloning and characterization of the warm-temperature-acclimation-associated protein Wap65 from carp, Cyprinus carpio

We determined full-length cDNA of carp warm-temperature-acclimation-associated 65-kDa protein (Wap65). It encoded 439 amino acid residues with a signal peptide of 22 residues and showed an amino acid sequence identity of 88% to that of goldfish reported before (J. Biol. Chem. 1995. 270: 17087–17092). The number of potential N-linked glycosylation sites of carp Wap65 was two in contrast to three for goldfish. In addition, molecular mass determined by SDS-PAGE was apparently different from that of goldfish. These results suggest that the amount of oligosaccharide is different between the carp and goldfish protein. As in goldfish, carp Wap65 mRNA showed marked accumulation in hepatopancreas of the 30 °C- acclimated fish, which was 8-fold higher than that of the 10 °C-acclimated fish. Carp Wap65 showed 30% amino acid identity to mammalian hemopexins, which appeared to be considerably low in comparison with those among mammalian hemopexins (72 to 80%), or among carp Wap65 and rainbow trout hemopexin-like protein (70%). However, although mammalian hemopexins contain residues comprising the heme binding pocket, carp Wap65 lacked one of the two histidine residues to serve as heme axial ligands in hemopexins. Our data on carp protein substantiates the previous observation for goldfish and indicates that Wap65 might have some important functions in warm-temperature-acclimation of fish.     

A critical consideration of the metamorphosis zone when identifying daily rings in otoliths of European eel, Anguilla anguilla (L.)

Otoliths of glass eels and larvae collected from the Rio Minho (Portugal/Spain) as well as from the Iberian continental slope from the Bay of Biscay were analyzed by scanning electron microscopy. Preliminary data are presented on the total radius of the otolith and the width of the zone exhibiting a diffuse structure which, in the literature, is suggested to be the zone of metamorphosis. It was found that the radius increased from the development stage I through stage II to the glass eel stage (Vb). The width of the diffuse zone also exhibited an increase. Calculations of the dimensions of the diffuse zone of specimens older than stage I revealed that the area of metamorphosis amounts to about 28–60% of the total diffuse zone. From these results it is evident that part of the diffuse zone must have formed during a larval phase of retarded growth, during which no formation of daily growth rings takes place. Only the outer portion of the diffuse zone can be accounted for by the metamorphosis. For these reasons, an exact age determination by counting daily rings seems impossible. A determination of the oceanic life of the eel recruits is difficult for other reasons too: all earlier and recent studies have indicated that Anguilla leptocephali and their metamorphosis stages do not occur on the continental shelf, which could add an additionally high amount of time needed until arrival at the coasts.     

Effects of dietary pyridoxine on disease resistance,immune responses and intestinal microflora in juvenile Jian carp (Cyprinus carpio var. Jian)

This experiment was conducted to evaluate the effects of dietary pyridoxine on disease resistance, immune responses and intestinal microflora of fish. A total of 1050 Jian carp (11.71 ± 0.05 g) were randomly distributed into seven groups, feeding diets containing graded levels of pyridoxine (0.2, 1.7, 3.2, 5.0, 6.3, 8.6 and 12.4 mg kg^?1 diet). After 80 days of feeding, a challenge trial was conducted by injection of Aeromonas hydrophila for 17 days. Results indicated that with increasing dietary pyridoxine concentration up to 5.0 mg kg^?1 diet, survival rate after challenge with A.hydrophila and phagocytic activity of leukocyte were improved (P < 0.05), and plateaued thereafter (P > 0.05). Red blood cell and white blood cell counts were lowest when fed the diet containing 1.7 mg pyridoxine kg^?1 diet. Haemagglutination titre, lysozyme activity, acid phosphatase activity, total iron‐binding capacity, antibody titre and immunoglobulin M content followed the similar pattern to that observed with survival rate. Aeromonas hydrophila, Escherichia coli and Lactobacillus counts in intestine were not affected by dietary pyridoxine concentration (P > 0.05). These results suggested that pyridoxine could enhance immune response of fish.     

Estimation of glass eel (Anguilla anguilla) exploitation in the Severn Estuary,England

Mark-release-recapture trials were conducted to determine the exploitation rate of glass eel Anguilla anguilla (L.) by handheld dip nets in the Severn Estuary in the spring of 2020. The glass eel were marked with Rhodamine B at a concentration of 0.1 g/L, 200 glass eel/L for 4.5 hr and then placed in fresh water for 36–48 hr to monitor mortality. In trial 1, 891 ± 100 of the 20,455 glass eel were recaptured while in trial 2,373 ± 172 of the 27,923 glass eel released were recaptured. The exploitation rate was estimated to be 4.36% (±0.49) in trial 1 and 1.33% (±0.62) in trial 2. The size of the glass eel population from trial 1 was estimated to be 24.69 t (22.46 – 28.81), and the overall exploitation rate of the fishery for the season was 7.8% (6.7% – 8.6%). Comparisons are made with studies in other estuaries and with conservation targets set by the EU Eel Regulation and the Eel Management Plan for the River Severn. The study suggests the fishery is not the main cause of the Severn RBD failing to meet escapement targets.     

Growth,body composition,intestinal enzyme activities and microflora of juvenile Jian carp (Cyprinus carpio var. Jian) fed graded levels of dietary phosphorus

A 9‐week feeding trial was carried out with juvenile Jian carp (Cyprinus carpio var. Jian) to study the effects of dietary phosphorus on growth, body composition, intestinal enzyme activities and microflora. Quadruple groups of juvenile Jian carp (7.17 ± 0.01 g) were fed practical diets containing available phosphorus 1.7 (unsupplemented control), 3.6, 5.5, 7.3, 9.2 and 11.0 g kg^?1 diet to satiation. Feed intake, specific growth ratio and feed efficiency were the lowest in fish fed the basal diet (P < 0.05). Body moisture, protein, lipid content and ash were all significantly affected by dietary available phosphorus levels (P < 0.05). Activities of trypsin, amylase, Na⁺, K⁺‐ATPase, alkaline phosphatase and gamma‐glutamyl transpeptidase were improved with increasing dietary phosphorus levels. Intestinal Aeromonas and Escherichia coli decreased with increasing dietary phosphorus up to 3.6 and 5.5 g kg^?1 diet respectively (P < 0.05), while Lactobacillus increased with the increasing dietary phosphorus up to 9.2 g kg^?1 diet (P < 0.05). These results suggested that phosphorus could enhance intestinal enzyme activities of juvenile Jian carp and the minimum dietary available phosphorus requirement for SGR of juvenile Jian carp (7.2–63.8 g) was 5.2 g kg^?1 diet.     

On the actual recruitment of European eel (Anguilla anguilla) in the River Ems,Germany

Data from the historical River Ems glass eel fisheries in the 20th Century have been used for the ICES recruitment series. Commercial catches ceased from 1980 and dropped to zero; thus, eel recruitment data are since lacking. The present study assessed the actual eel recruitment to the River Ems with regard to the local tidal conditions. Standardised measurements were performed on relative glass eel occurrence at the tidal weir from 2014 and quantitative sampling at the next weir upstream from 2013. Eel migration at the tidal weir was dominated by glass eels and increased 2–3 month earlier than at the upstream weir, where migration was dominated by pigmented eels. In 2016, glass eels were marked at the tidal weir and pigmented elvers were sampled at the upstream weir. The 2016 eel recruitment was estimated at about 1% of the mean historical recruitment. Eel recruitment at the two weirs was not directly linked. Many eels seem not to pass the upstream weir. Further investigations are needed to reveal whether the results represent a regular pattern. In the long run, the perpetuation of the recruitment series is also foreseen.     

Estimation of production and long‐term changes in the population of European eel Anguilla anguilla (L.) in an Irish lake

Monitoring of yellow‐phase and silver‐phase Anguilla anguilla during their continental life history is necessary for evaluation of stock recovery measures. Eel population data for an Irish lake (Lough Sheelin) were compiled for the period 1993–2014. Catch data from 2009 to 2014 provided minimum estimates of recent silver eel production ranging annually from 0.79 to 1.84 kg/ha. Long‐term changes in yellow eel abundance and silver eel size structure were assessed as part of a fishery monitoring programme. Yellow eel catch per unit effort (CPUE) in the lake (from longline surveys) was considerably higher in the 1990s (52.2–62.1 eels/100 hooks) than 2002–2008 (1.9–15.8 eels/100 hooks). Conversely, during 1993–2014, the mean size of silver eels migrating from the lake increased significantly (p < .001), from 659 mm to 838 mm. The results suggest that in the absence of direct yellow eel abundance data, interannual variation in silver eel size structure may be a useful monitoring tool for local eel stocks as part of Eel Management Plans (EMP's).     

鲤、鲇及草鱼肌肉理化特性的比较研究

以杂食性的鲤(Cyprinus carpio)、肉食性的鲇(Silurus asotus)和草食性的草鱼(Ctenopharyngodon idellus)为研究对象,测定了2龄鱼的背部、胸部、腹部和尾部肌肉的pH值、滴水损失、熟肉率、失水率、胶原蛋白含量及剪切值等理化指标,对3种鱼不同部位的肌肉理化特性进行了比较,初步探讨了鱼肉理化特性与食性的关系.结果表明,鲤、鲇及草鱼肌肉的pH值为6.2 ～6.7,3种鱼肌肉间pH值无显著差异(P＞0.05);鲤、鲇及草鱼各个部位肌肉之间以及3种鱼之间的肌肉滴水损失率均无显著差异(P＞0.05);每种鱼各部位肌肉之间的熟肉率无显著差异(P＞0.05),熟肉率表现为:草鱼＞鲤＞鲇(P＜0.05);同一种鱼不同部位肌肉的失水率无显著差异(P＞0.05),3种鱼之间的失水率表现为:鲇＞鲤＞草鱼(P＜0.05);同一种鱼不同部位肌肉的胶原蛋白差异显著(P＜0.05),3种鱼之间差异也显著,鲇＞鲤＞草鱼(P＜0.05);同一种鱼不同部位之间的剪切值差异显著(P＜0.05),3种鱼之间的剪切值表现为:鲇＞鲤＞草鱼(P＜0.05).综合比较可见,淡水经济鱼类的肌肉理化特性与其食性存在一定的相关性,肉食性鱼的嫩度偏低,剪切值大,胶原蛋白含量高,失水率和滴水损失较大,保水能力也相对较差,且同一种鱼各个部位间的肉质理化特性也存在一定差异.     

Immune response,disease resistance and intestinal microflora of juvenile Jian carp (Cyprinus carpio var. Jian) fed graded levels of pantothenic acid

This study was to investigate the effect of dietary pantothenic acid (PA) on the disease resistance, immune response and intestinal microflora on juvenile Jian carp (Cyprinus carpio var. Jian). Seven diets (4.0, 15.5, 25.6, 36.1, 45.9, 56.1 and 65.9 mg PA kg^?1) were fed to Jian carp (12.95 ± 0.03 g) for 9 weeks. After 9‐week feeding trial, the challenge experiment with Aeromonas hydrophila was conducted to determine the impact of PA on fish disease resistance. Survival rate after challenge was promoted with the increasing PA levels (P < 0.05). Blood counts also significantly increased up to the dietary PA level of 25.6 mg PA kg^?1 (P < 0.05). Leucocyte phagocytic activity, lectin potency, lysozyme and acid phosphatase activity, and total iron‐binding capacity were improved with increasing PA levels (P < 0.05). Serum immunoglobulin M level and agglutination antibody titre to A. hydrophila were increased (P < 0.05) in fish fed the diets with the dietary PA levels between 56.1 and 65.9 mg kg^?1. PA also promoted the growth and reproduction of Lactobacillus and depressed Escherichia coli and A. hydrophila (P < 0.05). These results suggested that pantothenic acid could improve disease resistance, immune response, and the balance of intestinal microflora in juvenile Jian carp.     

Escapement success and patterns of downstream migration of female silver eel Anguilla anguilla in the River Meuse

Abstract – Downstream migration of female silver eel Anguilla anguilla (L.) was studied by remote telemetry in the lower part of the River Meuse (Belgium and the Netherlands) using a combination of nine detection stations and manual tracking. N = 31 eels (L_T 64–90 cm) were implanted with active transponders and released in 2007 into the River Berwijn, a small Belgian tributary of the River Meuse, 326 km from the North Sea. From August 2007 till April 2008, 13 eels (42%) started their downstream migration and were detected at two or more stations. Mean migration speed was 0.62 m·s^?1 (or 53 km·day^?1). Only two eels (15%) arrived at the North Sea, the others being held up or killed at hydroelectric power stations, caught by fishermen or by predators or stopped their migration and settled in the river delta. A majority (58%) of the eels classified as potential migrants did not start their migration and settled in the River Berwijn or upper Meuse as verified by additional manual tracking.     

Assessing the impact of hydropower and fisheries on downstream migrating silver eel, Anguilla anguilla, by telemetry in the River Meuse

Abstract –  European eel Anguilla anguilla is in strong decline. We assessed the relative impact of fisheries and hydropower during the downstream migration of silver eel in the River Meuse in the Netherlands using radio-telemetry. For this, 16 fixed detection stations (Nedap Trail-System^®) were used, each covering the entire river width, including all outlets to sea, and two covering the entrances of the two hydropower stations present. In September 2002, 150 silver eels were surgically implanted with Nedap-transponders and released at the catch site. Of these, 121 started to migrate downstream of which 37% successfully reached the North Sea. Hydropower mortality was at least 9% and assessed to be 16–26%. Fisheries mortality was at least 16% (reported recaptures) and estimated to be 22–26%. A clear difference was found in passage behaviour at hydropower stations, where 40% showed recurrence, in contrast to the river stations where this hardly occurred, indicating a hesitation to pass the turbines. Also a difference was found in diurnal pattern; 63% of the eels that passed through the turbines did so during the first 5 h of the night, whereas for the stations on free-flowing sections this was only 35%.     

Inheritance and reliability of random amplified polymorphic DNA-markers in two consecutive generations of common carp (Cyprinus carpio L.)

Random amplified polymorphic DNA (RAPD) markers have been used in a variety of genetic studies in fisheries and aquaculture. Most population studies are performed without preliminary data demonstrating the Mendelian inheritance and reproducibility of RAPD markers. In this study, the inheritance and reproducibility of RAPD markers was examined in two consecutive generations of common carp, Cyprinus carpio L. Variability and segregation of RAPD markers were investigated in one F₁ progeny and three F₂ progenies. Seventy-four RAPD markers were generated by five primers using DNA extracted from the initial ornamental (koi) common carp female and wild-type colour common carp male. Fifty-five of these RAPD markers were transmitted to the F₁ progeny and the inheritance patterns were analysed. Twenty RAPD markers were fully reproducible and demonstrated dominant simple Mendelian inheritance patterns in two consecutive generations. Twenty-four RAPD markers were not reproducible in all progenies. Thirteen markers displayed inheritance ratios in the progenies that did not fit simple Mendelian inheritance patterns. Non-reproducibility of RAPD markers and distorted ratios may be caused by the absence of amplification, poor amplification or by the appearance of artefact bands. Random amplified polymorphic DNA markers with poor reproducibility and non-Mendelian inheritance can lead to misinterpretations of data in population studies, resulting in errors in the estimation of genetic diversity within and between individual populations. Therefore, it is recommended to first identify the set of reproducible RAPD markers that demonstrate Mendelian inheritance before application of the RAPD technique in population studies.