The systematic development and optimization of large-scale sperm cryopreservation in northern pike (Esox lucius)

In our study, a systematic development of a new large-scale sperm cryopreservation protocol was carried out in northern pike (Esox lucius). The effect of 2 sugar based (glucose and trehalose) extenders, 3 dilution ratios (1:3, 1:9 and 1:19) 2 vol straws (0.5 and 5 mL) and a 10 mL cryotube, 2 different cryopreservation methods (Polystyrene box-P. box and Controlled Rate Freezer-CRF), as well as 3 different thawing periods (3, 3.5 and 4 min) were investigated on the motility of thawed sperm. The glucose based extender showed significantly higher pMOT (1:3–18 ± 16%, 1:9–20 ± 13%, 1:19–16 ± 12%) at all dilution ratios than in the trehalose based extender (1:3–0.3 ± 1%, 1:9–1±1%, 1:19–4±2%). A similar tendency was recorded in VCL and STR at a ratio 1:3 and 1:9. No significant difference was measured in sperm movement between the P. box and CRF using the 0.5 mL straw. Similarly no significant difference was observed in all motility parameters with 10 mL cryotube frozen in CRF at a ratio 1:3–1:19. An effective and short thawing period (3 min) was experimentally specified for the 10 mL cryotube cryopreserved in the CRF. In all large-scale cryopreservation methods, high pMOT (straw CRF: 57 ± 10%, straw P. box: 50 ± 9%, cryotube CRF: 41 ± 10%), and STR were measured, and no significant difference was recorded in all motility parameters. Our results demonstrate the effectiveness of our newly developed extender and the applicability of 3 different large-scale cryopreservation methods in pike sperm. Our protocols could be new prospective candidates for future exploitation in hatchery practice.     

The reproductive success of pike, Esox lucius: aspects of fecundity, egg density and survival

Power-law relationships have been estimated between fecundity and fish length, weight and age for pike from two gravel pit lakes (Main Lake and St Peter's Lake) in Buckinghamshire, England. Relative fecundity, estimated at 17.6 eggs g⁻¹ for Main Lake pike and 19.4 g⁻¹ for St Peter's Lake pike, did not differ significantly between the lakes. Population fecundity for the Main Lake was estimated at 10.6 million eggs and 6.4 million eggs in 1986 and 1987, respectively.
Experimental work involving samples of eggs from 18 artificially fertilized Main Lake female pike showed a significant relationship between egg diameter and female length, and a significant difference between mean egg diameters for 41-cm and 101-cm females. No significant relationships were found between mean egg size and mean fry size, nor between fry length and adult length. Mean size differences between newly hatched, 18-day-old and 41-day-old fry from the 41-cm and 101-cm females were not significant.
Samples of eggs taken at 2-day intervals from 6 April to 5 May in a pike spawning area of the Main Lake revealed clumped distributions of eggs, with average egg densities ranging from 1 to 47 eggs per 0.071 m² (14–671 m ²) on sand and silt substrata. The maximum egg density of 51 per 0.071 m² (729 m⁻²) was found on flooded grass. The stages of development of the eggs were identified over the sampling period. Survival from stage 1 to fry was estimated at 3–6%. Egg losses were estimated at 9–10% day⁻¹.
Egg survival experiments gave overall hatching success rates of 11 % on sand/silt compared with 2.5% on aquatic plants (1986), and 18.9% on undisturbed sand/silt compared with 7.9% on disturbed sand/silt (1987). The effect of siltation on egg hatching is discussed.     

Fatty acid composition of wild and cultured northern pike (Esox lucius)

The aim of the study was to determine the effect of diet on the fatty acids composition of cultured northern pike (Esox lucius) fillet (mean body weight, 620 g; age, 2.5 years; commercial feed) profiles when compared with wild pike caught in Lake Dga? Wielki in northern Poland (mean body weight, 570 g; age 3.5 years). Cultured pike fillets contained several times more fat (2.40%) compared with wild fish (0.19%, P < 0.01). The total relative quantity of saturated fatty acids (SFA; % of total fatty acids) and unsaturated fatty acids (USFA) in cultured and wild pike were similar. Monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) content differed in wild and cultured fish. Cultured pike contained nearly two times more MUFA (30.90% compared with 15.97%; P < 0.01) but less PUFA (42.04% compared with 58.33%; P < 0.01), n‐3 PUFA (32.42% compared with 41.26%; P < 0.01), and n‐6 PUFA (6.47% compared with 15.79%; P < 0.01). The largest differences were noted with regard to docosahexaenoic acid (DHA) and arachidonic acid (AA), the levels of which in cultured and wild pike were 18.93% vs 27.96% and 0.73% vs 8.60%, respectively (P < 0.01). Quality compositions of the commercial feed and the cultured pike fillets were identical.     

Spatial genetic structure of northern pike (Esox lucius) in the Baltic Sea

The genetic relationships among 337 northern pike (Esox lucius) collected from the coastal zone of the central Baltic region and the Finnish islands of Aland were analysed using five microsatellite loci. Spatial structure was delineated using both traditional F-statistics and individually based approaches including spatial autocorrelation analysis. Our results indicate that the observed genotypic distribution is incompatible with that of a single, panmictic population. Isolation by distance appears important for shaping the genetic structure of pike in this region resulting in a largely continuous genetic change over the study area. Spatial autocorrelation analysis (Moran's I) of individual pairwise genotypic data show significant positive genetic correlation among pike collected within geographical distances of less than c. 100-150 km (genetic patch size). We suggest that the genetic patch size may be used as a preliminary basis for identifying management units for pike in the Baltic Sea.     

Genomic inbreeding and population structure of northern pike (Esox lucius) in Xinjiang,China

Northern pike (Esox lucius) was widely distributed in the high latitudes of the northern hemisphere. In China, northern pike was originally distributed only in the upper reaches of the Irtysh River in Xinjiang and has appeared in many water bodies outside the Irtysh River Basin in Northern Xinjiang. A total of four populations were collected from north to south in Xinjiang, including Irtysh River (RIR), Ulungu Lake (LUL), a small lake nearby Ulungu River (LJD), and Bosten Lake (LBO). We estimated population genomic parameters, performed gene flow analysis, and estimated the effective population size of each population. The proportion of individuals with high inbreeding coefficient (F ≥ 0.0625) accounted for 36.4% (44/121) of all sequenced individuals, approximately 4.5% (1/22) in LUL, 25.9% (7/27) in LBO, 42.9% (18/42) in RIR, and 60% (18/30) in LJD. RIR had the highest mean of genomic relatedness (coancestry coefficient = 0.025 ± 0.040, IBD = 0.036 ± 0.078). Gene flow results showed that the population spreading was from RIR into two branches, one was LBO, and the other continued to split into LUL and LJD, and migration signal from LBO to LUL was detected. Our results suggested that the extinction risk of northern pike was very low in Xinjiang of China, and the controlled capture fishery of northern pike could be developed reasonably.     

Mercury toxicity in livers of northern pike (Esox lucius) from Isle Royale, USA

Many laboratory studies have documented that mercury can be toxic to fish, but it is largely unknown if mercury is toxic to fish in their natural environments. The objective of our study was to investigate the toxic effects of mercury on northern pike (Esox lucius) at Isle Royale, Michigan. In 124 northern pike from eight inland lakes, concentrations of total mercury in skin-on fillets ranged from 0.069 to 0.622 microg/g wet mass (wet wt). Concentrations of total mercury in livers increased exponentially compared with concentrations in fillets, to a maximum of 3.1 microg/g wet wt. Methylmercury constituted a majority of the mercury in livers with total mercury concentrations <0.5 microg/g wet wt, but declined to 28-51% of the mercury in livers with total mercury concentrations >0.5 microg/g wet wt. Liver color (absorbance at 400 nm) varied among northern pike and was positively related to liver total mercury concentration. The pigment causing variation in liver color was identified as lipofuscin, which results from lipid peroxidation of membranous organelles. An analysis of covariance revealed lipofuscin accumulation was primarily associated with mercury exposure, and this association obscured any normal accumulation from aging. We also documented decreased lipid reserves in livers and poor condition factors of northern pike with high liver total mercury concentrations. Our results suggest (i) northern pike at Isle Royale are experiencing toxicity at concentrations of total mercury common for northern pike and other piscivorous fish elsewhere in North America and (ii) liver color may be useful for indicating mercury exposure and effects in northern pike at Isle Royale and possibly other aquatic ecosystems and other fish species.     

Mosaic individuals found in genetically manipulated northern pike (Esox lucius) using flow cytometry

The purpose of this study was to investigate abnormal larvae produced in the experimental induction of gynogenetic northern pike ( Esox lucius L.), using a flow cytometry method. We reported for the first time the mosaic individuals found in genetically manipulated northern pike. Two types of abnormal larvae were obtained after eggs were fertilized with UV-irradiated sperm and then exposed to heat shock treatment. One type had malformations frequently associated with haploid syndrome and were proven to be haploids. The other type had a similar body size and appearance as diploids but with curved body and a swirling swimming pattern. Individuals of the second type were proven to be mosaics containing both 1n (59.5 ± 14.7% mean ± SD) and 2n (29.7 ± 13.3%) cell populations. The ratio of 1n:2n cell populations ranged from 80.6:10.6 to 38.5:44.7. There was no significant difference in the relative nuclear DNA content and cell size between 1n and 2n cell populations of the mosaic individuals as compared to haploid cells and diploid cells, respectively. The nuclear DNA content of the northern pike was estimated to be 2.07 ± 0.06 pg ( n  = 10) given that rainbow trout has a red blood cell nucleus DNA content of 4.66 pg.     

The influence of angling-induced exercise on the carbohydrate metabolism of northern pike (Esox lucius L.)

Summary Capture by angling was used to induce burst exercise in northern pike. By 3 h after exercise blood lactate had risen to levels of 15.2 mmol l⁻¹ (Fig. 2), which greatly exceeded the maximum post-exercise levels (4.0 mmol l⁻¹) previously reported for muskellunge, a close relative of pike. White muscle lactate level was high, 41.8 mmol kg⁻¹, immediately after capture but declined to 23.2 mmol kg⁻¹ by 6 h (Fig. 2). Blood glucose level more than doubled after exercise and remained elevated even after 96 h of recovery (Fig. 2). During the first 6 h after angling, pike disposed of 9.57 mmol (861 mg) of lactate per kg body weight. A whole body metabolic rate of 153 mg O₂ kg⁻¹ h⁻¹ is sufficient to account for this rate of lactate removal through oxidation (Table 3). However, the metabolic rate of the highly oxidative organs and tissues (red muscle, gills, liver, kidney, heart, and spleen) must be very high (>1,000 mg O₂ kg⁻¹ h⁻¹) to oxidize even 60% of the lactate that disappeared from pike after exercise (Fig. 5). Mortality of pike from angling stress was less than 3%.     

Intensive fishing can mediate stronger size-dependent maternal effect in pike (Esox lucius)

In pike E. lucius L., evidence on maternal effect on reproductive output is mixed. We studied whether older and larger pike females produce eggs and larvae of higher quality (weight, starvation resistance) in three forest lakes in southern Finland. Later, the study lakes were subjected to intensive experimental pike fishing, which we assumed would increase resource availability and lead to higher maternal investment (larger egg size). Length of female pike was positively correlated with the dry weight of eggs and larvae but this relation was dependent on female age. In old females, the effect of female length on egg weight was lower or even negative. Survival analysis showed a positive effect of female length on larval survival time indicating that larvae from larger females are less vulnerable to starvation during the early stage of life. After the intensive pike fishing, the positive effect of female length on egg weight was stronger in all age classes probably due to the released resources. Based on the high quality and amount of reproductive products in large (but not very old) females, they are important for the reproduction of pike populations. This should be considered in fisheries management.     

Monoamine oxidase activity in embryos of pike (Esox lucius)

1. Mitochondrial MAO specific activity was measured in eggs and early embryos of the teleostean fish Esox lucius using tryptamine, 5-hydroxytryptamine (5-HT) and phenylethylamine (PEA) as substrates. 2. Tryptamine is the most readily deaminated substrate in mitochondria isolated from unfertilized eggs and embryos at the stages of cleavage, blastula and gastrula. 3. Monoamine oxidase activity gradually decreases during development and at the gastrula stage it is respectively 80% (tryptamine), 70% (5-HT) and 50% (PEA) of that found in the egg using the corresponding substrate. 4. The inhibition of egg MAO activity by clorgyline and deprenyl measured in E. lucius eggs using tryptamine as substrate, indicates the presence of a single form of MAO not corresponding to the MAO A and MAO B described in terrestrial vertebrates.     

Individual specialization and trophic adaptability of northern pike (Esox lucius): an isotope and dietary analysis

Northern pike (Esox lucius) are often considered to be specialist piscivores, but under some circumstances will continue to eat invertebrates as adults. To examine effects of fish assemblage composition on the trophic ecology of pike, we combined stable isotope analysis (SIA) of carbon and nitrogen and stomach content analysis (SCA) on pike from five lakes in northern Alberta, three of which contain only pike (“pike-only”) and two that also contain yellow perch (Perca flavescens) or white sucker (Catostomus commersoni) (“pike-other”). Fish were more important as prey and empty stomachs, which often characterize piscivores, were significantly more frequent in pike-other than in pike-only lakes. However, even though invertebrates were more important for pike in pike-only lakes, SIA and SCA indicated that invertebrates were also an important component of pike diets in pike-other lakes. SIA and SCA also revealed considerable intrapopulation variation in trophic ecology, with individuals in some populations differing by as much as two trophic levels. Comparisons of stomach contents and isotope signatures of the same fish suggested that within these variable populations, specialization on invertebrates or fish was a long-term trait of some individuals. SIA indicated that trophic position increased and diets shifted to a greater importance of littoral prey as pike grew in pike-only lakes, but not in lakes with other fish present. Trophic adaptability in northern pike is expressed at both the population level, where the trophic ecology is sensitive to differences in prey regimes, and at the organismal level, in the form of intrapopulation variation and individual specialization. Received: 1 July 1998 / Accepted: 3 February 1999     

Molecular cloning and sequence analysis of the cDNA for northern pike (Esox lucius) growth hormone.

The complete nucleotide sequence of the northern pike (Esox lucius) cDNA for pregrowth hormones was determined from clones derived from a pituitary gland cDNA library. Seventeen cDNA clones were isolated from a single mRNA species. A cDNA of 1,227 nucleotides was sequenced and found to encode a polypeptide of 209 amino acid residues, which included a putative signal sequence of 22 amino acid residues. Sequence comparison of the northern pike growth hormone gene to other known growth hormone genes revealed similarities closest to other members of the superorder Protacanthopterygii, which includes the Salmonidae family (i.e., salmon and trout).     

Seasonal dynamics of fatty acid composition in female northern pike (Esox lucius L.)

Seasonal changes in the fatty acid composition of neutral and polar lipids were measured in the ovary, liver, white muscle, and adipopancreatic tissue of northern pike. The role of environmental and physiological factors underlying these changes was evaluated. From late summer (August–September) to winter (January–March), the weight percentage of n-3 polyunsaturated fatty acids (especially 22:6n3) declined significantly in the neutral lipids of all somatic tissues examined. However, large quantities of n-3 polyunsaturated fatty acids accumulated in the recrude cing ovaries during fall and the weight percentage of n-3 polyunsaturated fatty acids in ovary polar lipids also increased significantly. Additionally, the n-3 polyunsaturated fatty acid content of somatic polar lipids increased significantly during fall due to increases in the total polar lipid content of the somatic tissues. This suggests that during fall n-3 polyunsaturated fatty acid are diverted away from somatic neutral lipids and thereby conserved for use in ovary construction and for incorporation into tissue polar lipids. The percentage of n-3 polyunsaturated fatty acid in ovary neutral lipids also declined during fall and early winter, perhaps as an adaptation to conserve these fatty acids for storage in oocyte polar lipids and later incorporation into cellular membranes of the developing embryo. Reductions in the n-3 polyunsaturated fatty acids content of somatic and ovarian neutral lipids during fall were compensated for specifically by increases in the percentage of monounsaturated fatty acids rather than saturated fatty acids. This suggests that the ratio of saturated to unsaturated fatty acids in pike neutral lipid, is regulated physiologically, and hence may influence the physiological functioning of these lipids. During fall and early winter the percentage of saturated fatty acids declined significantly in the polar lipids of all tissues examined. This change was consistent with the known effects of cold acclimation on the fatty acid composition of cellular membranes. As the ovaries were recrudescing from September to January, liver polar lipids exhibited significant decreases in the percentage of total polyunsaturated fatty acids and n-3 polyunsaturated fatty acids and increases in monounsaturated fatty acids, and acquired a fatty acid composition very similar to that of ovary polar lipids. Therefore, seasonal changes in the percentage of polyunsaturated and monounsaturated fatty acids in liver polar lipids probably reflect the liver's role in vitellogenesis rather than the effects of temperature on membrane fatty acid composition. At all times of year, the fatty acid compositions of white muscle and adipopancreatic tissue neutral lipids were very similar, which may indicate a close metabolic relationship between these lipid compartments.Abbreviations AP adipopancreatic - BHT butylated hydroxytoluene - CI confidence interval - EFA essential fatty acids - MUFA monounsaturated fatty acids - NL neutral lipids - PL polar lipids - PUFA polyunsaturated fatty acids - SFA saturated fatty acids     

The effects of extenders and sperm‐egg ratios on fertilizing ability of cryopreserved testicular sperm of northern pike (Esox lucius L.)

The effects of four different extenders and two sperm‐to‐egg ratios on fertilizing ability of cryopreserved testicular sperm of northern pike (Esox lucius L.) were tested in the present study. Testicular sperm was diluted with each of the four extenders (0.6 m sucrose + 15% DMSO, 0.3 m glucose + 15% DMSO, 0.6 m sucrose + 10% methanol and 0.3 m glucose + 10% methanol, all supplemented with 10% egg yolk and 1.7 g KCl L^?1) and frozen in 0.5‐ml straws at 2 cm above the surface of liquid nitrogen and thawed at 25°C for 30 s. Then 125 μl or 50 μl of frozen‐thawed testicular sperm was poured onto about 1250 eggs for fertilization. The results showed that both sperm‐to‐egg ratio and diluent had no significant influence on cryopreservation efficiency of testicular sperm, whereas cryoprotectant had a significant influence. The highest fertilization rate (92.2%) was obtained from testicular sperm cryopreserved in glucose‐based extender containing 10% methanol at a sperm‐to‐egg ratio of 1 × 10⁶ spermatozoa per egg. The results indicated that glucose‐based extender containing 10% methanol, 10% egg yolk and 1.7 g KCl L^?1 was a useful combination.     

The effect of invertebrate diets on lipids, fatty acid composition and physiological condition of pike larvae (Esox lucius)

The present study tests two invertebrate diets for pike larvae reared in floating cages. One diet contained mainly copepods and cladocerans such as Eucyclops serrulatus and Bosmina longirostris , while the other diet contained Eucyclops serrulatus together with a small proportion of large Chydoridae and chironomid larvae. During the first days of the experiment, the amount of food distributed to the larvae was insufficient (9–17.1 prey per larva and per day) and led to an increase in the mortality. Triacylglycerol contents of fry were low and dietary fatty acids were either catabolised or, concerning PUFA such as 22:6(n-3), incorporated into phospholipids. From day 13 to the end of the study (day 22), the most abundant diets distributed were accompanied by an increase in triacylglycerol PUFA and in triacylglycerol contents of larvae. During the same period weight and length growth were better for larvae reared on the copepod plus Chydoridae and chironomid diet, than for larvae reared on the copepod and Bosmina diet. The use of a lipid condition index based on the triacylglycerol/sterol ratio suggested that Chydoridae and chironomids positively influenced the growth and nutritional condition of larvae. The effects of prey type in terms of PUFA composition on pike larvae are discussed.     

Genetic characterization of 18 novel microsatellite loci in northern pike (Esox lucius L.)

The northern pike (Esox lucius L.), an important predatory freshwater species, is undergoing significant population decline. In this study, 18 novel polymorphic microsatellite loci were isolated and used for assessing genetic variation in the Chinese Ulungur and Hungarian Balaton populations of the species. The number of alleles ranged from 2 to 13, observed heterozygosity from 0.154 to 0.920 and expected heterozygosity from 0.145 to 0.921, thereby indicating the specific usefulness of these suites of markers for investigating genetic variability.     

Walleye, Stizostedion vitreum, and northern pike, Esox lucius, populations in three Alberta lakes

Results are presented from a survey of walleye and northern pike in Ethel, Marie and Wolf Lakes, Alberta conducted May-September, 1983. Three types of experimental gill net were fished, mainly inshore. Northern pike were abundant in all three lakes but white sucker and yellow perch also were common. Walleye were caught in largest numbers in Wolf Lake. Age and growth of both walleye and northern pike were determined from examination of the opercular bones. The walleye in the three lakes were represented by a few strong year classes but there was no synchronism between the lakes. The strong 1975 year class in Ethel and Wolf Lakes may have been correlated with high spring precipitation and heavy run-off. Although there were dominant year classes in the northern pike populations (especially the 1979 year class) the variability was not so marked as in the walleye. There were no significant differences between length-at-age for year classes of walleye in Ethel and Marie or northern pike in all three lakes. In Wolf Lake the 1978 and 1979 year classes grew faster than in previous years. Walleye and northern pike growth was well described by the von Bertalanffy growth model. Females of both species grew to a greater ultimate length and had a longer lifespan than the males. Fecundity data are presented for walleye from Wolf Lake (log₁₀ absolute fecundity = 0.856 + 2.441 log₁₀ length) and northern pike from Marie Lake (log₁₀ absolute fecundity =?2.671+4.052 log₁₀ length). The stomach contents of both species were examined during May and June. The majority of walleye had empty stomachs. Walleye and northern pike in Ethel and Wolf Lakes fed on a variety of fish and invertebrate species but both fed mainly on fish (in particular whitefish) in Marie Lake. The similarity of diet suggests competition for prey species. A walleye yield to commercial fishermen of c. 0.69 kg ha^?1 year^?1 in Wolf Lake has made up 8% of the total catch. Since 1970/71 when mesh size of nets was increased, the yield has been reduced to c. 0.30 kg ha^?1 year^?1. Anglers remove an additional c. 0.48 kg ha^?1 year^?1 walleye from Wolf Lake. Northern pike yields to commercial fishermen have been c. 2.00 kg ha^?1 year^?1 in Wolf Lake. Ethel and Marie Lakes have yielded only poor catches of walleye and northern pike. A yield model was used to illustrate that faster-growing northern pike have higher potential yields than walleye. Walleye produce higher yields in Wolf Lake than in Marie Lake, the reverse being true for northern pike. It is suggested that northern pike could be cropped at a higher rate in Marie and Wolf Lakes with a possible improvement in walleye stocks.