Induction of gynogenesis in muskellunge with irradiated sperm of yellow perch proves diploid muskellunge male homogamety

Diploid gynogenesis was induced in muskellunge Esox masquinongy using UV-irradiated muskellunge sperm as the first step in producing monosex females. In this approach, we have to rely on negative controls as an indirect reference for sperm genetic material destruction. In the first experiment, equal proportions of gynogenetic females and males were produced. Negative controls, UV-irradiated sperm without heat shock, yielded some normal hatching larvae, described as spontaneous diploids. In the second experiment, muskellunge eggs were activated using sperm from yellow perch. Because hybrids between these species are not viable, we produced unambiguous gynogens. When UV-irradiated yellow perch sperm was used to inseminate muskellunge eggs, haploids resulted (22.5% +/- 2.8% survival to the eyed stage). To produce diploid gynogens, a heat shock of 31 degrees C was applied to inseminated eggs 20 min after activation for a duration of 6 min. This process yielded several hundreds of gynogens for rearing. Several treatments of masculinizing hormone, 17alpha-methyltestosterone (MT), were carried out. Fish were dissected and gonads examined histologically for sex determination. Gynogens produced using yellow-perch sperm confirmed the presence of males in the control group, whereas the MT bath treatment (400 microg/liter) resulted in the production of fish with ovotestis. These results provide evidence for male homogamety in muskellunge and imply that a change of strategy is needed to produce monosex populations.     

Survival, growth and sex ratios of gynogenetic diploid honmoroko

Survival, growth and sex ratios of gynogenetic diploid honmoroko Gnathopogon caerulescens induced by blocking the release of the second polar body were examined. Mean survival of gynogenetic juveniles at 130 days after hatching was about 33% lower than that of the controls. No significant difference was seen in early growth between control and gynogenetic diploids. Standard length and body weight in six groups of gynogenetic progeny were significantly greater but in two groups were significantly smaller than in the controls. Although 69% of gynogenetic diploids had well-developed gonads, the remaining 30% had undeveloped gonads (small in size or thread-like), and those gonads were divided into four types. The mean proportion of females in the 10 gynogenetic groups was 87·2% which was significantly ( P <0·01) higher than in the controls (44·7%). Gynogenetic diploids included 3·0–35·3% males. Most of those males produced a high proportion of female progeny, but the proportion of male offspring varied widely. From these results, the sex determining mechanism in honmoroko was presumed to be female homogamety, but other factors resulted in the production of males.     

Masculinization of the gynogenetic juvenile ship sturgeon (Acipenser nudiventris Lovetsky, 1828) using 17α‐methyl testosterone

In sturgeons, the induction of gynogenesis and sex reversal could be important for potential production of neomale sturgeon and all‐female progeny for caviar production. The aim of this study was sex reversal of ship sturgeon (Acipenser nudiventris Lovetsky, 1828) gynogen into male sex. Five‐month‐old gynogens were sex reversed into male by including 17α‐methyl testosterone in their food for 7 months. Three treatments were considered as follows: (a) without treated (gynogen control), (b) 10 mg MT/kg diet, and (c) 50 mg MT/kg diet. All treatments (60 individuals) were sampled both the 30 and 36 months old and their sex was determined using classical histology method of gonad. The sex ratio of the progenies in the gynogen control were 73.3% female and 26.7% male. In treatment of 10 mg MT/kg feed, 18 specimens were studied that half of them (50%) showed pseudo‐testicular structure in the female gonad. That is 50% of the specimens were intersex, 27.7% were male and 22.3% were female. All of the fish fed by 50 mg MT/kg feed had been sex reversed to male. Sexual maturation of these fish had been recognized in stage III at 36 months old. In conclusion, 50 mg MT/kg feed found as effective dose for successful sex reversal in gynogenetic ship sturgeon.     

Steroid hormone profiles and correlative gonadal histological changes during natural sex reversal of sobaity kept in tanks and sea-cages

The gonadosomatic index of sobaity Sparidentex hasta fits well with the natural spawning season of February-March. At 12 and 24 months all the population matured as males. However, during this time, between April-September intersex gonads were seen. From October of the third year of life, females were seen for the first time. These changes in the histology of the gonads were correlated with the hormonal profile. Testosterone showed no significant correlation with sex but 11-ketotestosterone correlated very well with the mature male gonads and thus can be used for sexing. Oestradiol-17β higher in sex reversed individuals (females) in the spawning season than in males. The sequence of events in the protandrous sex change of this fish during the first two spawning seasons is male → intersex → male. However, during the third spawning season (age 36–37 months) this sequence is male →intersex → male or female (10–17%).     

Massive production of all-female diploids and triploids in the crucian carp

In many species of aquaculture importance, all-female and sterile populations possess superior productivity due to faster growth and a relatively homogenous size of individuals. However, the production of all-female and sterile fish in a large scale for aquaculture is a challenge in practice, because treatments necessary for gynogenesis induction usually cause massive embryonic and larval mortality, and the number of induced gynogens is too small for their direct use in aquaculture. Here we report the massive production of all-female triploid crucian carp by combining artificial gynogenesis, sex reversal and diploid-tetraploid hybridization. Previously, we have obtained an allotetraploid carp population (4n = 200) by hybridization between red crucian carp (Carassius auratus red var; ♀) and common carp (Cyprinus carpio; ♂). We induced all-female diploid gynogens of the Japanese crucian carp (Carassius cuvieri; 2n = 100). We also generated male diploid gynogens of the same species treated gynogenetic fry with 17-α-methyltestosterone, leading to the production of sex-revered gynogenetic males. Finally, these males were used to cross with the female diploid Japanese crucian carp gynogens and the allotetraploid females, resulting in the production of fertile all-female diploid Japanese crucian carp (2n=100) and sterile all-female triploid hybrids (3n = 150), respectively. Therefore, diploid crucian carp gynogenetic females and sex-reversed male together with an allotetraploid line provide an opportunity to produce all-female triploid populations in a large scale to meet demands in aquaculture industry.     

Gonadal morphology of female diploid gynogenetic and triploid rainbow trout

Chromosome sets of fishes can be manipulated; this practice includes the production of triploid and gynogenetic salmonids. Such chromosomal modifications often result in abnormal ovarian development. In rainbow trout (RBT), triploid females have string-like gonads lacking significant developing oocytes and are suggested to be sterile due to the odd set of chromosomes disrupting oogenesis. Aberrant ovarian development is reported to occur in about 30% of gynogenetic females. It has been suggested that gynogenetic fish are more prone to expressing developmental abnormalities due to either increased homozygosity or to incomplete inactivation of the paternal chromatin. This investigation was done to compare the ovarian morphology of female triploid and induced gynogenetic diploid RBT. The objective was to determine whether the presence of supernumerary chromosomal fragments, potentially generated during the process of sperm genome inactivation, would result in abnormal gonadal development in gynogens comparable to that observed in triploid females. Gonadal morphology was observed and karyotypical analysis was completed on 21 gynogenetic fish. In 90% of the fish examined, the presence of chromosomal fragments was positively correlated with irregular ovarian development. The atypical gonadal morphology observed in the gynogens resembled triploid RBT ovarian morphology. The results of this investigation support the hypothesis that disruption of the normal diploid chromosomal complement alters germ cell development in gynogenetic female RBT due to the unbalanced nature of the genome. J. Exp. Zool. 286:505-512, 2000.     

Growth and gonadal development of gynogenetic diploid Scophthalmus maximus

The survival, growth, gonadal development and sex ratios of gynogenetic diploid turbot Scophthalmus maximus between 9 and 36 months of age were investigated. Gynogenesis was induced by activation of freshly collected eggs with diluted (1 : 10 with Ringer) and UV‐irradiated (30000 erg mm⁻²) sperm, followed by a cold shock at −1 to 0° C for 25 min, starting at 6·5 min after fertilization. The exclusive maternal inheritance of the resulting gynogenetic diploids was confirmed in all individuals used in the experiment by microsatellite markers. Mean total length and mass throughout the experiment was higher in the controls than in the gynogenetics. The gonadal development in gynogenetic males proceeded normally, and in gynogenetic females it was delayed during the first sexual maturation but was normal during the second one. The sex ratio was 1 male (M) : 1 female (F) in the controls, as expected, but 1 M : 3 F in the gynogenetics. In a second, independent experiment, carried out with fish originating from different broodstock, however, the sex ratio was 1 M : 1 F in the controls and 0 M : 1 F ( i.e . 100% females) in the gynogenetics. Together, these results suggested that the main sex‐determining genetic component in turbot fits well to female homogamety, in accordance with data from other flatfishes. These results indicate that it will be possible to obtain near or all‐female turbot stocks through induced gynogenesis or by crosses involving hormonally sex‐reversed fish.     

Gonadal morphology of normal and sex-reversed triploid and gynogenetic diploid coho salmon (Oncorhynchus kisutch)

In this study, the gonadal morphology of untreated and sex-reversed juvenile triploid and gynogenetic diploid coho salmon was compared with that of diploids. Testes of triploids were of the same size as those of diploids. Spermatogonia, however, were significantly bigger than those of diptoids in both diameter (P<0·001) and volume (P<0·01), suggesting that this characteristic can be a useful indicator of ploidy in the early stages of gonadal development. In females, induction of triploidy did not affect the lamellar structure of the ovaries but reduced their size considerably. Further, these ovaries had no oocytes. Treatment of triploids with oestrogen resulted in the feminization of genotypic males, which had ovaries similar to those found in tripioid females. However, gonads of triploid males partially sex-re versed into females were identified by their enlargement, the presence of remnants of the male vascular system, and by the appearance of ovarian lacunae and germinal and somatic cells typical of triploid females, Induction of gynogenesis resulted in 100% females, of which 34% had ovaries of reduced size with areas devoid of oocytes. However, and contrary to what has been found in cyprinids, no male germ cells were observed in these ovaries. This discrepancy may reflect differences, in the mechanisms of sex determination between salmonids and cyprinids. Treatment of gynogenetics with androgen increased the number of fish with abnormal ovaries but also resulted in the production of phenotypic-male gynogenetic diploids, of which 11% had testes indistinguishable from those of untreated control diploids.     

Studies on the biology of reproduction in the cichlid Tilapia nilotica (L.): gonadal maturation and fecundity

Tilapia nilotica reaches maturity in the first year of life; females mature shortly before males (11·4 and 14·3 cm in length). The distinctive morphological features of the different stages of development of the gonads are characterized. The sex ratio varied in populations of different age (size)-groups: a ratio of 2·81:1 and 0·47:1 (females:males) was obtained in populations of young and old fish. In medium-sized populations the sexes were equally abundant. There is evidence that females breed more than once a season. Fecundity varied more with body length ( r =0·860) and weight ( r =0·806) than with age ( r =0·604). These variations were of the order of 2·02; 0·83 and 0·83 exponentials of length, weight and age, respectively.     

Depth distribution and biological characteristics of the European eel Anguilla anguilla in Lough Ennell, Ireland

Using a longline survey, a total of 196 European eels Anguilla anguilla were collected at different depths in Lough Ennell (maximum depth 30 m), central Ireland. The catch per unit of effort of A. anguilla that were caught from 1 to 25 m depths was lowest at 0·5–5·0 m and greatest at the deepest depth range (22·5–25·0 m). Sub-samples of A. anguilla from depths of <15 m showed little or no difference in size, sex ratio, age, growth rate, condition factor, length–mass relationship, gonado-somatic index, fin index or eye index with fish from depths of >15 m. All fish examined were female yellow-phase A. anguilla that had ages from 7 to 20 years (mean ± s . d . = 10·3 ± 2·9 years), with growth rates from 24·0–60·8 mm year⁻¹ (mean ± s . d . = 40·7 ± 8·5 mm year⁻¹). Variations in the growth rates were greater in the shallow group than that of the deep group. This study suggested that deeper regions are important feeding habitats for A. anguilla and that fish in this lake were growing moderately fast compared to similar habitats and areas in the species' range.     

Physiological impact of sea lice on swimming performance of Atlantic salmon

Atlantic salmon Salmo salar were infected with two levels of sea lice Lepeophtheirus salmonis (0·13 ± 0·02 and 0·02 ± 0·00 sea lice g⁻¹). Once sea lice became adults, the ventral aorta of each fish was fitted with a Doppler cuff to measure cardiac output ( ̇ ), heart rate ( f _H) and stroke volume ( V _S) during swimming. Critical swimming speeds ( U _crit) of fish with higher sea lice numbers [2·1 ± 0·1 BL (body lengths) s⁻¹] were significantly lower ( P  < 0·05) than fish with lower numbers (2·4 ± 0·1 BL s⁻¹) and controls (sham infected, 2·6 ± 0·1 BL s⁻¹). After swimming, chloride levels in fish with higher sea lice numbers (184·4 ± 11·3 mmol l⁻¹) increased significantly (54%) from levels at rest and were significantly higher than fish with fewer lice (142·0 ± 3·7 mmol l⁻¹) or control fish (159·5 ± 3·5 mmol l⁻¹). The f _H of fish with more lice was 9% slower than the other two groups at U _crit. This decrease resulted in ̇ not increasing from resting levels. Sublethal infection by sea lice compromised the overall fitness of Atlantic salmon. The level of sea lice infection used in the present study was lower than has previously been reported to be detrimental to wild Atlantic salmon.     

First report of intersex in a lutjanid: the goldband snapper Pristipomoides multidens

This is the first report of intersex in a lutjanid species, the goldband snapper Pristipomoides multidens, in which the gonads of a male fish contain multifocal oocytes scattered among testicular tissue. The incidence rate of intersex was low (<1.0%), with oocytes observed in the testes of only two of 206 male fish examined. The capacity for P. multidens to develop an intersex condition suggests that future monitoring of this species should include histological analysis of gonads.     

Short‐term freshwater exposure benefits sea lice‐infected Atlantic salmon

Atlantic salmon Salmo salar were infected with sea lice Lepeophtheirus salmonis (0·08 ± 0·007 sea lice g⁻¹) over a period of 4 h. Both infected and non‐infected fish were swim tested in sea water (SW) and fresh water (FW). The ventral aorta of each fish was fitted with a Doppler cuff in order to measure cardiac output, stroke volume and heart rate during swim testing. Blood samples were taken at rest and after exercise. Critical swimming speed of infected fish in SW (2·14 ± 0·08 body lengths, bl s⁻¹) was significantly lower ( P  < 0·05) than infected fish switched to FW (2·81 ± 0·08 bl s⁻¹) and non‐infected fish in SW (2·42 ± 0·04 bl s⁻¹) and FW (2·61 ± 0·08 bl s⁻¹). Cardiac and blood results indicated infected fish exposed to FW did experience stress, but relief from osmotic and ionic distress probably reduces energy expenditure, allowing the increase in performance. As the performance of sea lice‐infected fish improved upon transfer to FW, it is likely that heavily infected salmonids do return to FW to restore compromised osmotic and ionic balance, and remove sea lice in the process.     

Growth, gonadal development and sex ratios of meiogynogenetic diploid sea bass

Gynogenesis showed little effect on general physiology and gonadal development in sea bass Dicentrarchus labrax . Meiogynogenetic fish showed well-developed gonads indicating low occurrence of developmental imbalances even after gynogenesis induction in this species. In addition, the proportion of sexes of meiogynogenetic sea bass was similar to the diploid controls in two independent trials, which did not deviate significantly from a 1:1 male: female sex ratio. Even considering some environmental influence on sex differentiation, as has been previously demonstrated, the fact that the proportion of sexes was similar between gynogenetic and control diploids essentially eliminates the possibility that in the sea bass the females are the homogametic sex. Although the mechanism of sex determination of this species still remains unknown, even after gynogenesis induction, the genetic mechanism of the ZW/ZZ type could probably operate in the sea bass.     

Female-biased operational sex ratio of sexual host fish in a gynogenetic complex of Carassius auratus

To study the coexistence of sexual and gynogenetic forms, we examined the population structure of a gynogenetic complex of the Japanese crucian carp, Carassius auratus Temminck et Schlegel, during the April–June reproductive season by collecting 1225 mature fish that migrated from Lake Suwa to a tributary river for spawning. There were more sexual fish (about 80%) than gynogenetic fish in this complex, and the operational sex ratio in the sexual form was female biased (males were about 20%). Mean standard length and body weight of sexual females were larger than those of sexual males. Sex ratio was male biased in smaller fish (standard length, <8.5 cm) but female biased in larger fish (standard length, ≥8.5 cm). We determined age by scale ring marks; the average age of sexual females was higher than that of males, but there was no significant difference in the average age between sexual and gynogenetic females. Sex ratio in the sexual form was more female biased for old than for young fish, and the mean size of sexual females was larger than that of males of the same age. The clear female-biased sex ratio and age difference between sexual females and males can be explained either by (1) higher mortality of males or by (2) female-biased sex allocation. The latter process reduces the disadvantage of sex and contributes to the coexistence of sexual and gynogenetic forms. Received: November 24, 2000 / Accepted: March 6, 2001     

The biology of pike, Esox lucius L., in a southern eutrophic lake

The biology of pike, Esox lucius L., in a southern productive lowland lake is described. Scales and opercular bones were difficult to read and interpret, and growth estimated had perforce to be based on scale readings. Growth of Slapton pike is average and intermediate between recorded extremes. Both male and female pike reached maximum condition in February, just prior to spawning. The population of pike of over 450 mm fork length in the lake was estimated at 870·6 ± 389·3 in 1975, and 950·4 ± 143.3 in 1976/77. This leads to the conclusion that the pike population has one of the highest biomasses per surface area of water ever recorded. Number of pike per unit area of surface was also high, despite the fact that a significant proportion of the population (fish under 450 mm) could not be adequately sampled. Mean instantaneous mortality rate was 0·53, and mean instantaneous survival rate was 0·59. The roach provided the main item of diet of the pike, with perch taken less readily. Immature pike ate a significant proportion of invertebrates, but roach was again a common feature of the diet of even small fish. Two spawning migrations were identified; at other times of the year, pike were non territorial. The majority of pike spawned in March. Fecundity of a sample of females was assessed. Overall sex ratio was 1:1.     

Hypertension in Pagothenia borchgrevinki caused by X-cell disease

Approximately 15% of a population of the cryopelagic nototheniid fish Pagothenia borchgrevinki , found constantly swimming immediately beneath the annual fast ice, in McMudro Sound, Ross Sea, Antarctica, was affected by X-cell gill disease. This disease affected blood flow through the gill lamellae, and this in turn affected oxygen uptake. Exercise caused increases in heart rate and ventral aortic blood pressure. Heart rate increased from 15·1 ± 1·55 to 23·1 ± 0·93 beats min⁻¹ in healthy fish, with a similar increase (to 24·6 ± 0·26 beats min⁻¹) in X-cell-affected animals. In healthy fish, pressures rose with exercise (from 2·72 ± 0·11 to 3·75 ± 0·19 kPa) and then rapidly returned to resting levels during recovery. In X-cell fish pressures rose during exercise, but then continued to rise, to reach a high of 4·18 ± 0·13 kPa, close to the predicted maximum pressure able to be generated by these hearts. Recovery was rapid in healthy fish, but was prolonged in diseased animals. As they are constantly swimming, there is the potential that X-cell-affected fish suffer from chronic hypertension.     

Effects of angling on feeding by largemouth bass

Angling practice significantly effected the time required for largemouth bass Micropterus salmoides to begin feeding. Mean ± s.e. time until feeding resumed was longest for fish that experienced a simulated tournament (15·6 ± 2·2 h) followed by fish that were caught and released (8·4 ± 2·2 h) and controls (0·3 ± 1·6 h). Effects on feeding related to angling practices were maintained throughout the 48 h observation period. Using multiple logistic regression and bioenergetics simulations, decreased growth in fish subjected to competitive or catch-and-release angling events was predicted. Fish subjected to multiple captures in a pond experiment had greater mass loss than those not captured, supporting model predictions.     

Microhabitat selection in the early juvenile mudskipper Boleophthalmus pectinirostris (L.)

Using laboratory choice experiments, behavioural preferences of the early juvenile mudskipper Boleophthalmus pectinirostris for temperature, salinity and sediment were observed. The temperature preference experiments were conducted in an annular chamber with a thermal gradient of 27 to 34° C, and the fish selected a mean ± s . d . temperature of 31·2 ± 0·5° C. The salinity preference experiments were conducted in an aquarium with decreasing salinity from 20 to 15, 10, 5 and 0·5, and significantly more ( P < 0·05) fish were found in water with a salinity of 5. The sediment preference experiments were tested in circular tanks which contained four types of sediment (medium sand, fine sand, muddy sand and sandy mud), and the fish showed a clear preference for sandy mud. These results indicated that early juvenile B. pectinirostris showed behavioural preference for microhabitats. Temperature and salinity probably set large-scale boundaries on distribution, but sediment should be a critical factor for determining the distribution of the mudskipper.     

Nitrogen and phosphorus content of some temperate and tropical freshwater fishes

The nitrogen and phosphorus content of two temperate fishes, Rutilus rutilus and Perca fluviatilis , and six tropical fishes, Oreochromis niloticus , Cichla monoculus , Serrassalmus rhombeus , Plagioscion squamosissimus , Prochilodus brevis and Hoplias malabaricus , were investigated to test the hypothesis that variation in body P content and N:P ratio is related to body size. Regressions of %P and N:P ratios against fish size (length and mass) confirmed the hypothesis for P. fluviatilis and P. squamosissimus , suggesting that body size is an important factor driving body P content and N:P ratios in some fishes. Moreover, significant increases in %N and N:P ratio with body size was found for H. malabaricus , a common piscivorous fish of the Neotropics. Interspecific variation in %P and N:P ranged two-fold and significant differences ( P < 0·05) were found among the tested species. The mean ± s . d . elemental content across all fishes ( n = 170) was 10·35 ± 1·29% for N and 3·05 ± 0·82% for P, while the N:P ratio was 8·00 ± 2·14. Data on fish body nutrient content and ratio will improve parameterization of bioenergetics and mass balance models and help clarify the role of fishes in nutrient cycles in both temperate and tropical freshwaters.