Muscle fibre growth dynamics in diploid and triploid rainbow trout

The effect of triploidy on muscle fibre growth was determined by comparing hyperplasia and hypertrophy of white muscle fibres in all-female, diploid and triploid rainbow trout Oncorhynchus mykiss (100–400 mm total length). Conventional morphometry and protein and DNA concentrations were used to assess muscle fibre hyperplasia and hypertrophy in white muscle samples derived from an anterio-dorsal location. Muscle fibre distributions were significantly different between triploids and diploids in trout <300 mm. The proportion of fibres <20 μm was higher in diploids than in triploids and the proportion of fibres in the 20–40 μm category was higher in triploids than in diploids. This indicates that the hyperplastic fibres of triploids are larger than those of diploids. Larger hyperplastic fibres in triploids are probably due to the combined effect of increased nuclear size in triploids and the relatively high nucleus: cell ratio observed in small muscle fibres. These larger fibres may be less favourable to cellular metabolic exchange because of their smaller surface area to volume ratios, and perhaps account for reduced viability and growth observed in triploids during early life stages. On the other hand, the lack of difference in the distribution of fibres <20 μm between diploids and triploids at larger body size ranges (301–400 mm) imply that triploid trout may have higher rates of new fibre recruitment and growth capacity at these sizes. There was no difference between diploid and triploid trout in the mean size of muscle fibres; however, the number of fibres per unit area was reduced by 10% in triploids. No differences were observed in protein or DNA concentrations in muscle tissues between the two genetic groups. Since triploid nuclei have 1·5 times more DNA than diploid nuclei, this deviation from the expected muscle DNA concentration (1·3–1·4 times more DNA in triploids when the 10% reduction in fibre density is considered) suggests that the number of nuclei per muscle fibre is reduced. In both diploids and triploids, mean fibre size increased with body length while fibre density decreased. Similarly, protein concentration in the muscle tissue increased and DNA concentration declined with increasing body length. Protein/DNA ratio was strongly and positively correlated with fibre size. These results demonstrate that changes in DNA and protein concentrations can be used to assess hyperplasia and hypertrophy in muscle tissues. However, the morphometric procedure provides better insight into muscle fibre growth as it enables the direct visualization and analysis of muscle fibre distribution patterns.     

The critical swimming velocity of diploid and triploid brook trout

No significant difference was found in the critical swimming velocity of diploid and triploid brook trout Salvelinus fontinalis , suggesting that the triploid state is not associated with a reduced aerobic capacity.     

Effects of exogenous estradiol-17β on early growth and gonadal development of diploid and triploid female rainbow trout (Oncorhyncus mykiss)

Triploidy is a viable condition in teleosts. However, in many salmonids, the triploid condition in the female results in sterility as gametogenesis appears to be disrupted. Although the underlying mechanisms regulating the gonadal development of teleosts have not been clearly elucidated, the reversal of phenotypic sex by the administration of the appropriate exogenous steroid during early development supports the argument that gonadal steroids play a pivotal role in sexual differentiation and subsequent gonad development in these fish. To determine whether the failure of normal ovarian development in triploid female rainbow trout (Oncorhynchus mykiss) is due to an absence or reduction of endogenous sex steroids, ovarian morphology was compared between diploid and triploid juvenile rainbow trout treated with exogenous estradiol-17β (E₂). The ovaries of both untreated and E₂ treated diploid fish, at 145 days post-fertilization, contained synchronously developing oocytes in the perinucleolar stage, whereas ovaries from untreated and estradiol-treated triploid fish of the same age were considerably smaller and devoid of developing oocytes. No differences in the ovaries of triploid untreated fish and triploid fish treated with E₂ were observed. It is reported that exposure to exogenous E₂ during the period of gonadal differentiation is not sufficient to induce oocyte development in triploid rainbow trout. © 1996 Wiley-Liss, Inc.     

The feeding response of diploid and triploid Atlantic salmon and brook trout

Pair and group experiments were conducted to determine whether differences exist in feeding success between juvenile diploid and triploid salmonids in a competitive situation. In the pair experiments, 22 pairs (one diploid and one triploid) of size-matched Quebec-strain brook trout Salvelinus fontinalis (7·2–46·3 g) were fed an unlimited number of pellets three times a day for 5 days. Dominance was assigned to the fish which ate the most pellets within each pair. In the group experiments, groups of three diploid and three triploid size-matched fish were fed a restricted ration three times a day for 5 days. Hierarchical rank within the group was assigned based on the number of pellets consumed by each fish. The group experiment was repeated 10 times with Atlantic salmon Salmo salar (5·1–62·7 g), Quebec-strain brook trout (11·8-110·8 g), and large UNB-strain brook trout (18·2–33·0 g), and 12 times with smaller UNB-strain brook trout (0·6–2·0 g). A statistically significant difference in rank between ploidies was found only for the smaller UNB-strain brook trout in the group experiments, with diploids dominant over triploids. This suggests that there may be a difference in competitive feeding success between diploid and triploid brook trout early in development, but that this difference diminishes as the fish grow.     

Comparative growth performance of diploid and triploid European sea bass over the first four spawning seasons

During their 3–4 first years of life, triploid sea bass Dicentrarchus labrax grew in a similar fashion to diploids in fork length but more slowly than diploids ( P <0·05) in body weight, even when the diploids reached full sexual maturity. However, from 48–53 months of age triploids exhibited non-significantly higher instantaneous growth rates, and thus when fish were 4 years or older, differences in weight with diploids were no longer apparent, suggesting that triploidy could be of benefit in the culture of large (>1 kg) sea bass. The condition factor was reduced in both ploidies during the spawning season which took place in winter when the temperature was low. These observations suggest that any growth advantage in triploids, which were functionally sterile, may be offset by unfavourable environmental conditions. Thus, the potential gain of triploid fish, because they do not direct energy to gonadal growth, could not overcome the effects of low temperature on somatic growth, which coincided with the spawning season. This suggests that the low growth of this species during winter is more a consequence of low temperature than of the energetic cost associated with reproduction. On the other hand, the lower hepatosomatic index in triploid females in contrast to diploid females might be indicative of the lack of gonadal oestradiol-mediated hepatic synthesis of vitellogenin. Also, erythrocyte and haematocrit measurements showed an increased nuclear and cellular volume in triploids, but with similar cell numbers to those of diploids, respectively.     

Comparison of reproductive development in triploid and diploid female rainbow trout Oncorhynchus mykiss

The diploid rainbow trout Oncorhynchus mykiss reached sexual maturity 3 years after hatching and its oogenesis underwent four stages, which were oogonia, primary oocyte, secondary oocyte and egg. Reproductive development and hormone changes of 4 to 35 month‐old female O. mykiss were investigated using histological and radioimmunoassay methods in order to provide a theoretical and practical basis for the use of triploid female O. mykiss. The oogonium of the triploid female could develop into the oocytes of the prophase with abortion occurring later; the oogonium was surrounded by stroma cells to form the oogonium cluster and the gonads showed a virilescent tendency when the oogonium clusters were gradually replaced by spermatogenic‐like cytocysts. After 13 months, amounts of gonadotropic hormone (GtH‐I, GtH‐II) and oestradiol (17β‐E₂) in triploid females were lower than in diploid fish at corresponding time periods, but the amounts of testosterone (T) increased consistently after 21 months and were more than in diploid fish in the corresponding time periods (P > 0·05). The infertility of triploid females resulted from meiosis failure, which caused developmental abortion of oocytes and oogonium formed cytocysts before the prophase oocytes. The cytocyst formation was due to the lack of the normal interaction of ovum and follicular cells, the development of follicular cells producing steroids were inhibited, the arylate path from T to 17β‐E₂ was interrupted, concentration of 17β‐E₂ decreased and concentration of T increased in the blood, the content of vitellogenin (Vg) decreased in the liver with a low 17β‐E₂ and high T caused to ovaries to show a tendency to be virilescent.     

Influence of growth rate on white muscle dynamics in rainbow trout and brook trout

In trout, fast growth stimulated white muscle fibre hypertrophy ( P 0·001) and hyperplasia ( P <0·01) in outer fibres but not in deep fibres. Glycogen was most prevalent in outer fibres ( P <0·01) and in brook trout ( P <0·01) that on average had three to four times larger fibres than rainbow trout.     

Growth dynamics of white and red muscle fibres in fast- and slow-growing strains of rainbow trout

Compared with fish of a slow-growing strain, fast-growing rainbow trout exhibited significantly smaller white fibre diameters, throughout development from hatching to 24 cm body length, although possessing similar total number of fibres. In contrast, in red muscle, no differences were observed in fibre diameter between the two strains, but the fast growing fish showed a significantly higher number of red fibres. The differences in growth rate between the two strains were related to the mean white fibre diameter and were found to be matched by proportional adjustments in recruitment of new fibres to the growing muscle. Thus, the largest and fastestgrowing strain showed evidence of sustained higher recruitment of muscle fibres that endowed this strain with the potential to maintain rapid somatic growth for longer and accomplish greater muscle growth.     

RNA/DNA ratio as a growth indicator of stream periphyton

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Juvenile growth and aggression in diploid and triploid Chinook salmon Oncorhynchus tshawytscha (Walbaum)

Multilocus heterozygosity, aggressive and feeding behaviour, plasma cortisol levels and growth rate were evaluated among three groups of juvenile Chinook salmon Oncorhynchus tshawytscha : diploid, triploid and mixed groups of diploid and triploid fish. There was no difference between diploid and triploid fish in measurements of heterozygosity calculated using seven microsatellite loci, and these measurements did not correlate with performance measurements including feeding rate and growth rate. Aggression trials that examined small groups of fish revealed that after 4 days together in tanks, triploid fish were significantly less aggressive during feeding than diploid fish or fish in mixed groups. At the end of the trials, however, plasma cortisol levels did not differ among the three groups. Thirty-day growth trials in duplicate tanks of 60 fish revealed no difference in growth rate among diploid, triploid and mixed groups, but plasma cortisol levels were significantly lower in triploid fish than in either diploid fish or the mixed fish. Overall, independent of the above differences in aggressive behaviour and cortisol levels, these results suggest similar performance in diploid and triploid Chinook salmon, and thus provide support for the viability of triploid Chinook salmon culture in commercial aquaculture.     

Effects of progesterone and estradiol on the reproductive axis in immature diploid and triploid rainbow trout

In fish species, many studies demonstrated the crucial role of estradiol (E2) in the development of the reproductive axis, but progesterone (P) has been described mainly as a precursor steroid and no clear role by itself has been reported. Moreover, a cooperative effect of P (or another progestin) and E2 in fish has never been reported to our knowledge. In the present work, we investigated the effects of P, alone or in combination with E2, on the reproductive-axis of immature rainbow trout (Oncorhynchus mykiss). Liver vitellogenin and estradiol receptor (rtER) mRNA levels increased after E2 treatment, but were unchanged by P treatments as a reflection of peripheral action of steroids. In contrast, at the pituitary level, LH contents increased after E2 and/or P treatments. Focusing on the brain level, we confirmed a clear up regulation of rtER expression by E2 in sterile triploid females, and we also demonstrated a similar stimulating effect of P alone but no cooperative effect together with E2. In conclusion, our data demonstrate that in immature trout, prior to the beginning of the first reproductive cycle, unlike E2, P is able to stimulate the reproductive brain-pituitary axis without affecting vitellogenin synthesis in the liver.     

Biochemical indices as correlates of recent growth in juvenile green turtles (Chelonia mydas)

Nucleic acid and protein concentrations and their ratios are increasingly used as correlates of nutritional condition and growth in marine species. However, their application in studies of reptile growth has not yet been validated. The green turtle (Chelonia mydas) is an endangered marine reptile for which assessing population health requires knowledge of demographic parameters such as individual growth rates. The purpose of this study was to evaluate a number of biochemical indices ([DNA], [RNA], RNA:DNA ratio, [protein], protein:DNA ratio, and RNA:protein ratio) in liver, heart, and blood as potential predictors of recent growth rate in juvenile green turtles under controlled feeding conditions. Intake of juvenile green turtles was manipulated over twelve weeks to obtain a range of growth rates. With the exception of [RNA]_blood, [DNA]_heart, and [protein]:[DNA]_liver, all biochemical indices demonstrated significant linear relationships with growth rate during the last 1.5 weeks of the study. The best single predictors of recent growth were hepatic [RNA] and [RNA]:[protein], which explained 66% and 49%, respectively, of the variance in growth. Contrary to expectations, these two indices were negatively correlated with growth rate. To investigate the possibility that hepatic [RNA] was higher in slow-growing turtles because of elevated expression of antioxidant genes, we quantified glutathione peroxidase activity and total antioxidant potential. Both measures of antioxidant function were affected by intake and growth histories, but these effects did not explain our results for hepatic RNA and protein concentrations. We developed a model that predicted 68% of the variance in specific growth rate (SGR) with the equation SGR = − 0.913(ln[RNA]_liver) + 17.689(Condition Index) + 4.316. In addition, our findings that [DNA] and [RNA]:[DNA] for blood were significantly correlated with SGR demonstrate the potential utility of minimally invasive tissue sampling that could facilitate instantaneous population monitoring.     

Do random fluctuations in the intervals between feeding affect growth rate in juvenile three-spined sticklebacks?

This study measured the effects of regular and irregular intervals between feeds on the growth performance of juvenile Gasterosteus aculeatus . The experimental period was 21 days at 14) C and photoperiod of 10L: 14D. The fish were housed individually. The control fish received a constant ration every day. Fish on a constant interval were fed on days 1, 5, 9, 13, 17 and 21. Fish on a random regime were fed with the same average interval between feedings, but the interval varied randomly between 1 and 5 days. The rations were calculated so that over the 21-day period, all fish were supplied with the same total quantity of food. The two ration levels were: 2% (maintenance ration) and 6% of the initial body weight per day. At a given ration, the feeding interval had no significant effect on specific growth rate, RNA/DNA ratio and lipid contents. The percentage dry matter was slightly, but significantly lower in treatment groups than in the control group. Groups receiving a mean 2% ratio consumed all the food supplied. At a mean ration of 6%, the control group ate 100%, the regular interval group 95·4% and the irregular interval 98·3% of the total food supplied. For the temporal patterns of feeding used, the fish were able to adjust their food intake, when food become available, to compensate for short periods of food deprivation and maintain their growth performance except for a decrease in dry matter content.     

Deviation from morphological intermediacy in interstrain hybrids of rainbow trout,Salmo gairdneri

Synopsis Deviations from morphological intermediacy in six first generation hybrids between three hatchery strains of rainbow trout, raised in a common environment, are reported. Hybrids have higher mean counts of four meristic characters than their maternal parental strain in a significantly greater number of cases (18 out of 24). Furthermore, eight of eleven hybrid indices are not intermediate. These results are discussed in reference to several mechanisms and models proposed to account for observed responses of meristic characters to environmental and genetic influences.     

Control of skeletal muscle fibres and adipose cells size in the flesh of rainbow trout

The distributions of the diameters of skeletal muscle fibres and adipocytes were studied in rainbow trout. The cellularity of perivisceral adipose tissues and subcutaneous ventral and dorsal adipose tissues were characterized more specifically. In these tissues, a population of small adipocytes was distinguishable from larger adipocytes. The same was observed in white muscle. The effects of extrinsic factors (dietary lipid in two different thermal conditions) and intrinsic factors (strains in two different saline conditions, growth hormone) on the long-term response of the cellularity of both muscle and adipose tissues were studied. The effects of thermal environment were tested on fish fed the same ration and the effects of saline environment on fish fed ad libitum. The mean size of white muscle fibres was relatively unaffected by the different treatments tested: genetic origin and dietary lipid in different environmental conditions. There were significant differences in growth rate due to genetic origin and saline environment. The possible involvement of hyperplasia in response to these different factors is discussed. Growth hormone supplementation enhanced the percentage of small diameter fibres indicating a role of this hormone in the control of muscle hyperplastic growth. The mean size of adipose cells was affected only slightly by the different treatments tested. An increase in adipose cell size with aging and lipid content was observed. The percentage of small adipocytes also increased with aging. Thus, it is proposed that the development of adipose tissues, and thus fat retention, both result from the recruitment of new adipocytes and from the increase in size of existing adipocytes. The hyperplastic process contributed significantly to the differences in fat retention due to different treatments tested (strains, thermal and saline environments). When partially substituting fish oils for corn oils in the diet, a large increase in the ventral adipose cell size was seen indicating a potential negative effect of n-6 fatty acids on cell proliferation. Growth hormone treatment, on the contrary, induced a decrease in the size of perivisceral adipocytes. Thus, diet and hormonal status affect adipose cells size through two different metabolic pathways: lipogenesis and lipolysis respectively.     

Nutritional condition, starvation status and growth of early juvenile Japanese sea bass (Lateolabrax japonicus) related to prey distribution and feeding in the nursery ground

The nutritional condition and protein growth rates of Japanese temperate bass larvae and juveniles were studied in relation to prey distribution and feeding habits in the nursery grounds in Chikugo estuary, Ariake Sea, Japan. Samples were collected from a wide spatial area covering the nursery grounds of the fish in March and April 2003. Food habits of the fish were analyzed by examining the gut contents. Fish condition was evaluated by using RNA/DNA ratio and other nucleic acid-based indices and protein growth rates. The nucleic acid contents in individually frozen larvae and juveniles were quantified by fluorometric method. Two distinguished feeding patterns, determined by the distribution of prey copepods, were identified along the nursery ground. The first pattern showed the dependency of the fish on the calanoid copepod Sinocalanus sinensis, which was the single dominant prey in low saline upper river areas and the second pattern involved a multi-species dietary habit mainly dominated by Acartia omorii, Oithona davisae and Paracalanus parvus. Values of RNA, DNA, total protein, growth rates and for all the nucleic acid-based indices were higher in upstream areas than in the downstream areas. The proportion of the starving fish was higher in the downstream areas than in the upstream areas. Condition of juvenile sea bass was not equal throughout the nursery grounds; fish in the upper river were in better condition than those in the lower estuary. We speculated that utilization of S. sinensis, which appears a suitable prey item and provide a better foraging environment in the upstream nursery ground, is one of the key factors for early survival and growth of Japanese temperate bass larvae and juveniles in the Chikugo estuary.     

Comparison of growth performance,biochemical and fatty acid compositions between all‐female diploid and triploid rainbow trout Oncorhynchus mykiss (Walbaum, 1792)

This study compares the performance (in terms of survival and growth), biochemical and fatty acid compositions among all female diploid (AFD) and triploid (AFT) rainbow trout (Oncorhynchus mykiss). AFD and AFT fish with mean initial weights of 1,040.1 ± 1.3 and 1,039.7 ± 1.6 g, respectively, were reared and fed from March to August with a commercially extruded trout feed in a commercial freshwater fishfarm located in the Black Sea region (Samsun, Turkey). Survival was reduced throughout the experimental period in the AFT group with increasing water temperatures. At the end of the experiment, survival rates were 98.57 ± 1.43% and 82.38 ± 7.39% for the AFD and AFT groups, respectively. The AFD group showed significantly better growth performances in terms of weight gain, feed conversion rate (FCR), relative growth rate (RGR) and specific growth rate (SGR) than the AFT group (p < .05). Significantly less protein and greater fat content were also observed in the AFT group (p < .05). There was no significant difference between groups for fatty acid composition in meat, except for stearic acid (SA, C18:0) and docosahexaenoic acid (DHA, 22:6 n‐3). While the greatest SA concentration was in the AFT group, the greatest DHA concentration was observed in the AFD group (p < .05). The results indicate that female triploid rainbow trout are more susceptible to suboptimal environmental conditions (especially to higher water temperatures) than female diploids. Although poor triploid performance was observed in this study, relative productivity might be enhanced by rearing triploids in optimal environmental conditions.     

Nucleic acids in ichthyoplankton ecology: a review, with emphasis on recent advances for new perspectives

Nucleic acid assessments and especially the RNA/DNA ratio are used widely as indices of growth rate or nutritional condition of larval fish. Methodological aspects are considered with the aim of defining a most reliable analytical procedure. Laboratory calibrations can give conflicting results, but general and common features may be drawn from a global examination. A high variability of RNA content, and thus RNA/DNA ratio, was revealed by estimates at the individual level and seems to be more related to feeding condition in late larval or juvenile stages than in yolk-sac and first-feeding larvae. An alternative is suggested, based on the relative DNA content which appears more stable and sensitive to starvation during these early stages. The consistency of such a pattern is strongly supported by findings about regulation of ribosomal RNA content of tissues, and especially white muscle, by nutrition. Determination of a threshold founded on a simple observation of morphological development stages is proposed as a base to define the best way to assess the larval nutritional condition. Some field applications are reviewed briefly. Finally an appraisal of the advantages and drawbacks of the different nucleic acid-based indices is attempted, which permits delineation of some prospects for future research.