Kurokura Solution as Immobilizing Medium for Spermatozoa of Tench (Tinca tinca L.)

This study tested KUROKURA solution (Kurokura et al., 1984, Aquaculture 37, 267–274) and its modifications (by increasing NaCl content to 160, 180 and 200 mM) on immobilizing properties for sampling and short-term preservation of potential motility of tench spermatozoa. The immobilizing solution is used because, when collected, the sperm of most samples is contaminated by urine, causing spermatozoa to be of poor quality, with low motilities and velocities (almost 0), thus resulting in a worsened fertilization and hatching rate. Sperm was sampled with a syringe containing an immobilizing solution (IS), allowing an IS:sperm ratio of 2:1, under aerobic conditions at 0–4°C. This sperm solution was stored for 10 h and untreated sperm was collected prior to fertilization as a control. Spermatozoa quality was evaluated for the cell motility and velocity parameters and also for fertilization ability and hatching rate. Results obtained for tench sperm motility, velocity, fertilization and hatching rate showed that only sperm collected in the various immobilizing solutions can be successfully used for artificial insemination and preservation after 10 h at 0–4°C. The best immobilizing solution was found to be KUROKURA 180 (180 mM NaCl, 2.68 mM KCl, 1.36 mM CaCl₂· 2H₂O and 2.38 mM NaHCO₃), giving a fertility and hatching rate of 41%, with no change in rates after 10 h storage of sperm. Control sperm without immobilizing solution showed a fertilization and hatching rate of only 6–7%.     

Insemination, fertilization and gamete management in tench, Tinca tinca (L.)

Various procedures for artificial insemination in tench, Tinca tinca (L.) were re-examined with evaluation of fecundity of males and females among different tench strains. The objectives of this study were to enhance fertilization and hatching rates through optimization of the activation solution, the insemination process, the activation of gametes, and the elimination of eggs stickiness. Sperm for all experiments was collected directly into immobilization solution of modified Kurokura solution containing 180 mM of NaCl and stored at 2 °C for 2.5–5 h prior to the experiment. When dechlorinated tap water was used for activation a gamete ratio of 1150 spermatozoa per egg showed the best significant fertilisation and hatching rates. Optimal ratio between eggs (weight in g) and activation solution (in cm³) was 1:1. Different concentrations of activation solutions such as NaCl from 0 to 68 mM (0–136 mOsmol kg⁻¹) without buffer statistically decreased fertilization and hatching rates. The activation solution containing 17 mM of NaCl, 10 mM Tris–HCl, pH 8 and 9 significantly increased fertilization and hatching rates compared to dechlorinated tap water of pH 7 or activation solution containing 17 mM of NaCl, 10 mM Tris–HCl, pH 6 and 7. Adhesiveness of the eggs was successfully removed by incubation in Alcalase and activity: 3.16 Anson units per cm³.     

Effects of different diets on juvenile tench, Tinca tinca (L.) reared under controlled conditions

In a laboratory experiment at 28 °C, four commercial starters (Carp Starter, Uni Starter, Perla Plus, Asta) were fed intensively to 11-month-old juvenile tench Tinca tinca of initially 41 mm TL and 0.70 g BW, over a period of 120 days. Frozen chironomid larvae were fed to fish below satiation as a reference diet for comparison of the fish body chemical composition. Final survival rates of 99–100% and satisfactory growth were attained with all dry diets, ranging from 73.3 (Carp Starter) to 84.8 mm TL (Asta) and from 5.39 (Carp Starter) to 8.09 g BW (Perla Plus). External body deformities of up to 96.4% specimens appeared in those fed Uni Starter, Carp Starter and Perla Plus, whereas 0% were recorded for Asta and chironomids. Fish in groups with deformities showed significantly elevated lipid content in their bodies, lower ash content and higher condition coefficient values. Juveniles in groups without deformities did not substantially change lipid content, but the ash content increased. Thus, among four dry diets studied, only Asta starter can be fed effectively and safely to juvenile tench reared intensively under controlled conditions.     

Studies on sperm of diploid and triploid tench, Tinca tinca (L.)

The tench Tinca tinca is an interesting fish from the viewpoint of polyploidy and related atypical reproduction aspects. Triploid tench were produced artificially. Studies of spermiation as well as of sperm motility and structure were performed on several triploid and diploid males simultaneously with individual experimental crosses with diploid females to define their reproductive capacities. The testes of triploids visually looked less developed in the most of cases with lower sperm production (0.05 cm³ sperm per male), GSI and weight of testes compared to diploids (0.58 cm³ sperm per male). Analysis of variance showed significant influence of ploidy level on the percentage of motile spermatozoa. Triploidy did not change percentage of live spermatozoa and velocity of spermatozoa at the first time of sperm movement. The study of sperm structure by scanning electron microscopy revealed that most sperm cells were of normal structure with some anomalies. Sperm heads of triploid and diploid males were mostly round-shaped, 1.86±0.2 and 1.6±0.18 μm in diameter. The midpiece of triploid spermatozoa was slightly narrower than that of diploid ones with typical cylindrical shape. Flow cytometry revealed sperm cells of triploids to be largely aneuploid (1.47 n) with high mosaic DNA, oscillating from haploid DNA content (1.0 n) to diploid DNA content (1.9 n). Experimental crosses between triploid males and diploid females revealed that these males were capable to stimulate effective development with relatively high level of fertilization and hatching rates from 0 to 70%. In conclusion, triploidization does not seem to guarantee sterility of tench.     

Influence of nutrition on the lipid quality of pond fish: common carp (Cyprinus carpio) and tench (Tinca tinca)

Like marine fish freshwater fish are an important source of essential fatty acids for human nutrition. However, the fatty acid composition of pond fish can vary considerably and strongly depends on that of the ingested food. Investigations on the fatty acid composition of common carp (Cyprinus carpio) and tench (Tinca tinca) have shown that different methods of rearing and feeding cause substantial variations in the proportions of the n-6 and n-3 polyunsaturated fatty acids of these fish species. Carp reared on the basis of natural food in ponds exhibit high contents of n-6 as well as n-3 fatty acids in their muscle triacylglycerols. On the other hand carp fed supplementary wheat in ponds resulted in somewhat lower levels of these essential fatty acids. High amounts of n-3 fatty acids can be found in carp fed high-energy diets containing high levels of fish oil. Analogous results were obtained in experiments with tench reared under different nutritional conditions. While rearing on the basis of only natural food in ponds as well as feeding supplementary wheat yielded in similar levels of n-3 and n-6 polyunsaturated fatty acids, higher contents of n-3 fatty acids were recorded in tench fed pellets. High levels of n-3 polyunsaturated fatty acids in foodstuffs have positive effects on human health. Experiments with different cultured fish species proved that the fatty acid composition of the edible parts can be influenced by the diet. Therefore, a finishing diet with a suitable fatty acid profile can be used to improve the nutritional quality of fish products of farmed origin.     

Selective behaviour of a tench, Tinca tinca (L.), stock in different light–substrate combined conditions

Two physical factors light or substrate (fibre glass, artificial vegetation, gravel and mud) were determined in a 24 m long artificial channel to have importance on the distribution of 33 tench 2+, mean size 11.8±0.9 cm S.L. For this purpose in five trials tench could select between the half of the channel occupied by their preferred light intensity (inferior to 10 lux) combined with the less preferred substrate (fibre glass) or the half with the combination of one of the substrates considered and higher light conditions (40 lux). Fish preferred, with a 100% frequency, the optimal light-less preferred substrate when compared with the combinations fibre glass, gravel and mud. When artificial vegetation was the substrate considered, tench selected the combination preferred substrate – non-optimal light intensity with a 98.26±2.68% frequency. This result changed when increasing light intensity up to 150 lux, avoiding the shading effect. We demonstrated that light has a bigger influence than substrate in juvenile tench habitat selection under controlled conditions.     

Quantity, motility and fertility of tench Tinca tinca (L.) sperm in relation to LHRH analogue and carp pituitary treatments

The spermiation of tench males was stimulated with Supergestran containing mammalian LHRHa lecireline at the following doses: 5, 10, 20 and 40 g kg⁻¹ b.w.; then with carp pituitary suspension (CPS) at a dose of 2 mg kg⁻¹ b.w. and with a control of saline physiological solution. The following days, meaning 24, 48 and 72 h after injection, sperm was collected to evaluate volume and the number of sperm per male per kg body weight (B.W.) The percentage of motile sperm and velocity of spermatozoa were measured 48 h after hormonal injection, and 72 h after hormonal injection the sperm was evaluated for fertilization and hatching ability. All 42 males in experimental groups were diploid. Live weight did not differ significantly among experimental groups. The strongest stimulation of spermiation was achieved with LHRHa in dosage of 20 and 40 g kg⁻¹ b.w. and CPS compared to males of the control group and lower dosage of LHRHa. Analysis of variance showed no significant influence of the treatment on the velocity and percentage of motile spermatozoa. The effect of different treatment on the fertilization capacity (the number of spermatozoa per egg was equilibrated) was significant. Significantly the highest quality of sperm collected 72 h after injection expressed by percentage of fertilization and hatching (62–65% fertilization and 61–64% hatching rates, respectively) was found for LHRHa in dosage of 20 and 40 g kg⁻¹ b.w. Significantly the lowest parameters of fertilization and hatching were found for the control group, on the 12% level.     

Morphological changes of digestive structures in starved tench Tinca tinca (L.) juveniles

The aim of the study was to evaluate the effect of short-term starvation on the morphology of the digestive system of juvenile tench Tinca tinca (L.). The morphological changes in digestive structures of juveniles, fed 70 days starter or Chironomidae larvae, were examined after 1, 12, 24 and 48 h of starvation at 28 °C. The morphological changes in the organisation of the digestive structures were as follows: (1) Intestine – proteolysis of the intestinal mucosa, especially of enterocytes in apical part of mucosal folds; (2) Liver – progressive reduction of lipid vacuoles and glycogen granules, and, in effect, volume reduction of hepatocytes cytoplasm; (3) Pancreas – progressive degeneration of exocrine pancreatic cells, parallel with decrease of zymogen activity. The fish fed natural food prior to starvation seem to be more resistant to starvation than those fed starter.     

Long-term examination of health conditions in monoculture of communally tested amphimictic diploid, diploid gynogenic and triploid tench, Tinca tinca (L.)

Amphimictic diploid (2n), induced triploid (3n) and meiotic gynogenic (MeiG) tench populations were established in 1998 by mass propagation. After separate nursing, fish were group-marked by freeze branding and 200 specimens per population were stocked into an experimental earthen pond for communal test until present. Parasitological and haematological (haemoglobin content Hb, haematocrit PCV and differential leukocyte count) examinations were performed every spring and autumn in the 2+ category, once sex could be registered macroscopically. Altogether 9 genera and/or species of parasites were detected on skin, 7 genera and/or species on gills, 1 genus in eye and 1 species in gut. The highest invasion extensity was recorded in the MeiG tench in the majority of examinations till the age of 4+. The courses of PCV and Hb values for 2n and MeiG populations showed high similarity while those for 3n population were slightly decreased and sex-related differences proved to be insignificant within populations in the majority of examinations. Differential leukocyte counts showed significant differences (p<0.05) of 3n vs. 2n and/or MeiG considering lower proportions of lymphocytes and higher proportions of segmented neutrophils in the majority of cases.     

Changes in commercially exploited populations of tench, Tinca tinca (L.), in littoral zones of lakes of northeastern Poland

Commercially exploited tench, Tinca tinca, (L.) populations in 480 lakes in the northeastern Poland were analyzed on the basis of the fishery data from 1949 to 1994. The long-term changes in tench catch were studied in four categories of lakes with respective proportions of littoral zones (<30, 31–60, 61–90 and <90% of total lake area). The analyses were performed with using relative catch (kg ha⁻¹ month⁻¹) of tench. Mean relative catch of tench for 1949–1994 ranged from 0.36 kg ha⁻¹ month⁻¹ in lakes with smallest littoral zone (>30%) to 0.93 kg ha⁻¹ month⁻¹ in lakes with the largest littoral (<90%). Mean relative catch increased with increasing proportions of littoral zones in lakes. However, long-term trends in mean relative catch of tench showed decreasing tendencies in all lakes. The beginning of these tendencies was found about ten years earlier in lakes with smaller littoral (<60%) in comparison to lakes with larger littoral zones (>60%). Mean stocking rates of tench (number of 1-year-old fish per ha of littoral) for 1949–1994 were not significantly different in lakes of studied categories. However, lakes with proportionally less littoral zone had reduced stocking efficiency estimated as number of stocked fish per ha of littoral to require 1 kg ha⁻¹ month⁻¹ of tench.     

The effect of feeding on oxygen consumption and ammonia excretion of juvenile tench Tinca tinca (L.) reared in a water recirculating system

The effect of feeding frequency (one, three, and continuous feeding), feed ration (0.2, 0.5, 0.8% of total fish biomass), and feeding per se on the oxygen consumption (OC, mg O₂ kg⁻¹ h⁻¹) and ammonia excretion (AE, mg TAN kg⁻¹ h⁻¹) of juvenile tench (body weight 15–19 g) and variations in these parameters in daily cycles were examined. Fish metabolism was studied in a recirculating system (rearing tanks of 0.2 m³, water temperature 23 °C). It was found that oxygen consumption and ammonia excretion depended significantly on feed ration. An increase of feed ration from 0.2 to 0.8% of fish biomass caused an increase of OC and AE from 126.80 mg O₂ kg⁻¹ h⁻¹ and 1.95 mg TAN kg⁻¹ h⁻¹ to 187.35 mg O₂ kg⁻¹ h⁻¹ and 8.80 mg TAN kg⁻¹ h⁻¹ (p<0.05). There was no dependence between feeding frequency and the mean rate of oxygen consumption. However, the relationship between feeding frequency and ammonia excretion by juvenile tench was statistically significant (p<0.05). Feeding frequency significantly affected daily fluctuations of AE and OC. It was found that diurnal variations in metabolic rates were strictly related to tench feeding, and the daily variations of AE were significantly higher than OC.     

Tench (<Emphasis Type="Italic">Tinca tinca</Emphasis> L.) larvae rearing under controlled conditions: density and basic supply of <Emphasis Type="Italic">Artemia</Emphasis> nauplii as the sole food

After artificial reproduction of tench, larvae must be maintained indoors, and studies on rearing conditions are needed, focussing on the reduction of labour and costs. Three experiments on larvae (5th day post-hatch) were conducted for 25 days using Artemia nauplii as the sole food in order to determine basic feeding and density conditions during the first rearing period. Tench were maintained in 25 l fibreglass tanks, supplied with an artesian water flow throughout of 0.2 l min⁻¹. Water temperature was 22.5 ± 1°C, and the photoperiod was natural. Larvae fed on a restricted amount of nauplii reached high survival rates, even with the minimum of 50 nauplii larva⁻¹ day⁻¹. This amount of food may be sufficient at least for the first 25 days of exogenous feeding if fast growth is not the priority, and high densities can be maintained with good survival rates (over 90% up to 160 larvae l⁻¹ and 77% with 320 larvae l⁻¹). When food was supplied in excess once a day, high survival rates were achieved (91–97%), without differences among the densities tested. Animals at a density of 100 l⁻¹ reached the highest length (15.57 mm) and individual weight (46.8 mg). This growth is greater than those reported in studies feeding several times a day. It could be deduced that, while live food remains available for tench, it is not necessary to feed so frequently. Considering the relationship among the initial number of animals, final survival and growth and ration supplied, the new data reported here are useful to establish suitable stocking densities under both culture and experimental conditions.     

池塘两种养殖方式下草鱼的营养差异

为了解传统池塘养殖和池塘内循环系统养殖草鱼的营养差异,对2种养殖方式草鱼的营养成分及营养品质进行了比较研究。结果显示,池塘养殖草鱼肌肉水分、粗脂肪含量、脏体比以及pH降低值和滴水损失高于循环养殖系统的草鱼,而肌肉粗蛋白含量和抗氧化能力低于循环养殖系统的草鱼。池塘养殖和循环系统养殖草鱼肌肉必需氨基酸构成比例均符合FAO/WHO的标准,必需氨基酸均是赖氨酸(Lys)含量最高;池塘养殖草鱼氨基酸总量(∑TAA)、鲜味氨基酸总量(∑DAA)和甜味氨基酸总量(∑SAA)低于循环养殖系统;根据氨基酸评分(AAS)和化学评分(CS),2种模式的第一限制性氨基酸均为缬氨酸(Val)。循环养殖系统草鱼肌肉∑n-3PUFA和∑n-6PUFA含量显著低于池塘养殖,但肌肉EPA+DHA含量是池塘养殖的1.5倍。研究表明,草鱼营养组成合理,池塘内循环系统养殖的草鱼在营养成分及鲜甜味方面优于池塘养殖。     

龙须菜新品系GL 家系F1 配子体的性状分析

本研究旨在为深入构建龙须菜(Gracilaria/Gracilariopsis lemaneiformis)育种体系提供基础数据。采用DTOPSIS综合评价法对实验室构建的龙须菜GL家系的F1配子体的性状进行分析与评价,以期把F1配子体的综合性状这一模糊量化指标转化为该株系对理想解的相对接近度Ci值的量化指标,进而对各藻株的综合性状优劣进行综合分析,为未来良种的杂交以及选育提供依据。所测的100株藻体中,线性生长速率变化范围为2.0~7.3%/d,变异系数为26.79%,标准差达到1.28,各藻株之间存在明显的差异。藻株的分枝数分布很不集中,变异极为显著,其平均数为1.93个,标准差为0.8,变异系数为41.17%,差异明显,变异幅度相对很大。主枝基部直径变化范围1.4~2.2 mm,标准差达到0.18,变异系数在3个性状中最小,为9.87%。线性生长速率、分枝数、基部直径等性状分布均呈现明显的正态分布特征,标准差分析显示,各性状相对本身参数变化幅度较大,说明F1配子体存在一定的性状变异,可以挑选性状差异大的藻种进行杂交以期获得优良藻种。DTOPSIS综合评价分析显示20、47、93、6、31株系综合性状的评价值比较高,为后期人工选育优良藻种提供依据。     

Detection of hybrids between masu salmon <Emphasis Type="Italic">Oncorhynchus masou masou</Emphasis> and amago salmon <Emphasis Type="Italic">O. m. ishikawae</Emphasis> occurred in the Jinzu River using a random amplified polymorphic DNA technique

Identification of masu salmon Oncorhynchus masou masou and amago salmon O. m. ishikawae was accomplished using a random amplified polymorphic DNA (RAPD) technique. Of 80 primers screened, three primers amplified subspecies-specific fragments (OPA11-1095 ^*, OPB5-1618^* and OPD5-2038 ^*). Based on fragment patterns, 33 hybrids were detected among 150 upstream-migrating individuals in the Jinzu River, central Japan, a long-time habitat of masu salmon, to which amago salmon were recently introduced. All of the individuals examined in the 2000–2002 cohorts were identified as F ₁ and F _x as well as pure masu and amago forms. These results indicated continuing hybridization, the genetic pollution of the native masu salmon population by amago salmon possibly being a serious problem in the Jinzu River. Both standard length and body weight in F ₁ hybrids tended to be less than in pure masu salmon. However, F _x hybrids showed similar body sizes to masu salmon.     

Neuroimmunological regulation of α-MSH release in tilapia (Oreochromis mossambicus)

This study describes the effects of IL-1 (interleukin 1) and LPS (bacterial endotoxin lipopolysaccharide) on the release of α-MSH (alpha melanocyte stimulating hormone) from the neurointermediate lobe (NIL) of the teleost Oreochromis mossambicus (tilapia). In vivo treatment of tilapia with IL-1 for 8 days led to a 49% inhibition of basal α-MSH release, measured by means of an in vitro micro-superfusion technique. The treatment did not affect the sensitivity of the tissue to TRH. In vitro, the release of α-MSH was inhibited by LPS in a dose dependent manner. In addition to its effects on the unstimulated release of the hormone, LPS also blunted the response to a TRH stimulation. Together with recent results obtained by others demonstrating the effects of (neuro-)peptides on immune parameters and the presence of cytokines in fish, the present data establish the bidirectional character of the communication between the immune and the (neuro-)endocrine systems in teleosts.

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Résumé Cette étude décrit les effets de l'IL-1 (interleukin 1) et du LPS (bacterial endotoxin lipopolysaccharide) sur la libération de α-MSH (alpha melanocyte stimulating hormone) par le lobe neurointermédiaire (NIL) d'un téléosteen Oreochromis mossambicus (tilapia). Le traitement in vivo du tilapia avec l'IL-1 pendent 8 jours conduit à une inhibition de 49% de la libération basale d'α-MSH mesurée à l'aide d'une technique in vitro de micro-superfusion. Ce traitement ne modifie pas la sensibilité du tissue au TRH. In vitro, la libération de α-MSH est inhibée par le LPS de manière dose-dépendante. En plus de ses effets sur la libération basale de α-MSH, le LPS bloque aussi la réponse à une stimulation par le TRH. Confrontés à des donnés récemment publiés montrant les effets de (neuro-)peptides sur les paramêtres immunitaires et la présence de cytokines chez les poissons, nos résultats établissent le caractère bidirectionnel de la communication entre le système immunitaire et les systèmes (neuro-)endocriniens.

     

Effects of different food items on the culture of the mysid shrimp Mysidopsis almyra (Crustacea: Pericaridea) in a static water system

The effects of several food items on larvae production and survival ofthe mysid Mysidopsis almyra were compared. A total of sixdiets were used. The diets were: 1) phytoplankton (Isochrysisgalbana), 2) an artificial diet (Liqualife®, Cargill,Minneapolis, MN), 3) a mixed diet composed of both zooplankton (mostlycopepods)and phytoplankton, 4) 750 mg g^–1 of HUFA enrichedArtemia nauplii and 250 mgg^–1 of the artificial diet, 5) newly hatchedArtemia nauplii (24-hour incubation at 28°C) and 6) newly hatched Artemia naupliienriched with HUFA (SELCO®, INVE Inc., Ghent, Belgium) for 12 hours. Mysidsfed HUFA enriched Artemia nauplii (diet 6) had the highestproduction and survival rates, although not significant (P > 0.05), comparedto diets 3, 4 and 5, while the phytoplankton and the artificial diet hadsignificantly lower production and survival rates (p > 0.05).     

Studies on digestibility and digestive enzyme activities in Labeo rohita (Hamilton) juveniles: effect of microbial α-amylase supplementation in non-gelatinized or gelatinized corn-based diet at two protein levels

A 60-days feeding trial was conducted to delineate the effect of both gelatinized and non-gelatinized corn with or without supplementation with exogenous α-amylase at two dietary protein levels (35% and 28%) on dry matter digestibility, digestive enzymes and tissue glycogen content of Labeo rohita juveniles. Three hundred and sixty juveniles (average weight 10±0.15 g] were randomly distributed into 12 treatment groups with each of two replicates. Twelve semi-purified diets containing either 35% or 28% crude protein were prepared by including gelatinized (G) or non-gelatinized (NG) corn as carbohydrate source with different level of microbial α-amylase (0, 50, 100 and 150 mg kg⁻¹). The dry matter digestibility of G corn fed groups was significantly higher (P < 0.05) than that of the NG corn fed groups. Hepatosomatic index (HSI), liver glycogen and intestinal amylase activity of G starch fed groups were significantly higher (P < 0.05) than those of the NG corn fed groups. However, the reverse trend was found for gastrosomatic index (GSI), muscle glycogen and intestinal protease activity. Addition of 50 mg α-amylase kg⁻¹ feed improved the dry matter digestibility of NG starch fed groups, which was similar to that of the G corn fed groups or NG corn supplemented with 100/150 mg α-amylase kg⁻¹ feed. HSI, liver glycogen and intestinal amylase activity were significantly increased (P < 0.05) at minimum level of α-amylase in the feed (50 mg kg⁻¹) and did not increase due to further inclusion of amylase in the diet. Supplementation with α-amylase at 50 mg kg⁻¹ increased the intestinal amylase activity beyond which no significant changes were observed. Protease activity of liver and intestine was highest (P < 0.05) in higher crude protein (CP) fed groups, but protease activity of the intestine was significantly higher in the α-amylase supplemented groups. Hence, it was concluded that feed with 28% CP containing either G corn without α-amylase or NG corn with 50 mg α-amylase kg⁻¹ may be used as the alternative carbohydrate source for L. rohita juveniles.