Manchurian Trout Research Articles

Effects of Exogenous Hormones on Spawning Performances, Serum Gonadotropin and Sex Steroid Hormone in Manchurian Trout (Brachymystax lenok) during Sexual Maturation

The objective of this study was to investigate the effects of salmon gonadotropin releasing hormone analog (S-GnRH-A) and human chorionic gonadotropin (HCG) on the serum levels of gonadotropins (GtHs) and sex steroid hormones in Manchurian trout during sexual maturity. Females in the treatment group were injected with 20 µg/kg S-GnRH-A and 400 IU/kg HCG, whilst males were injected with half the amount. Blood samples were collected at 0, 5, 10, and 20 days post injection (dpi). The results showed an increase in female follicle-stimulating hormone (FSH) levels in the treatment group at 5 dpi, and from 5 dpi onwards, a difference was observed between the groups, with higher FSH levels in the treatment group compared to the control group. In males, FSH levels showed group differences at only 5 dpi (p < 0.05) and were higher in the treatment group. In females, luteinizing hormone (LH) peaked at 10 dpi in the treatment group, and there were differences between the two groups at 10 dpi and 20 dpi. In males, LH was significantly higher in the treatment group than in the control group at 5 dpi and 10 dpi (p < 0.05). In females, estradiol (E2) was significantly higher in the treatment group than in the control group at 5 dpi and 10 dpi (p < 0.05), whereas in males, the difference between the two groups was only observed at 5 dpi (p < 0.05). The female testosterone (T) level in the treatment group was significantly higher than that of the control group at 5 dpi and 10 dpi (p < 0.05), while T levels were significantly higher in the male treatment group than in the control group at 5 dpi (p < 0.05). The level of 17α–hydroxyprogesterone (17α-OHP) in the females of the treatment group was significantly higher than that of the control group at 5 dpi and 10 dpi (p < 0.05), and 17α-OHP levels were significantly higher in the male treatment group than in the control group at 10 and 20 dpi (p < 0.05). The fish were successfully spawned after S-GnRH-A and HCG injections, and brood amount, fertilization rate, and hatching rate were significantly increased (p < 0.05). This study provides a framework for a better understanding of the mechanisms of exogenous hormone-mediated control of reproduction in Manchurian trout.

Effect of Different Feeding Rates for the Growth and Body Composition of Manchurian Trout, Brachymystax lenok

A 50-day feeding trial was conducted to investigate the effects of feeding rates (FRs) on the growth, feed utilisation, and body composition of Brachymystax lenok. Juveniles of B. lenok (initial body weight of 7.6 ± 0.2 g) were fed a commercial diet at the different FRs of 1, 2, 3, 4, and 5% of their body weight. The results indicated that weight gain (WG) and specific growth rate (SGR) were first increased and then decreased with an increase in the FR. The WG and SGR of the 4% group were 299.78% and 3.96%, respectively. The feed conversion ratio (FCR) and protein efficiency ratio (PER) were also significantly affected by the FR ( P < 0.05 ). Furthermore, the condition factor (CF) did not differ significantly among the treatment groups ( P > 0.05 ). However, the FR significantly affected the hepatosomatic index (HSI) and visceral somatic index (VSI) ( P < 0.05 ). With an increase in the FR, the moisture content of the fish decreased, but the ash content changed irregularly. The protein content increased significantly with increasing FR ( P < 0.05 ). The lipid content showed a trend similar to protein in the body. Based on the SGR and FCR data, the optimal FR range was 2.4 to 3.1% for B. lenok (7−30 g body weight) at a water temperature of 16°C.

The relationship between the population characteristics and physical habitat of Manchurian trout (<i>Brachymystax lenok tsinlingensis</i>) in the Geybangcheon stream

The relationship between the population characteristics and physical habitat of Manchurian trout (<i>Brachymystax lenok tsinlingensis</i>) in the Geybangcheon stream

Effects of dietary LNA/LA ratios on growth performance, tissue fatty acid composition and immune indices in Manchurian trout,Brachymystax lenok

This study aimed to assess the effects of different linolenic acid (LNA, 18:3n-3) and linoleic acid (LA, 18:2n-6) ratios in diet on growth performance, fatty acid composition in muscle and liver and non-specific immunity in serum, liver and intestines of juvenile Manchurian trout (Brachymystax lenok). With casein, soybean meal and corn protein meal as main protein sources, a vegetable oil-based (VO-based) diet with iso-nitrogen (40%) and iso-lipid (18.5%) was prepared by adding different proportions of linseed oil, corn oil and palm oil. The LNA/LA ratio in the VO-based diets was 0.02, 0.55, 1.06, 1.61 and 2.17 (constant composition of LNA and LA). The control group (CON) used fish oil (FO) as the sole lipid source. Healthy juvenile Manchurian trout (body weight: 7.00 ± 0.07 g) were fed in a recirculating aquaculture system for 9 weeks. The results showed that VO-based diets did not affect the growth of Manchurian trout, but decreased DHA and EPA levels in the dorsal muscle of Manchurian trout. Levels of LA, arachidonic acid (ARA) and n-6 polyunsaturated fatty acids (PUFA) of liver in VO-based diets were significantly higher than those in CON diet and decreased with an increasing LNA/LA ratio in the diet. The composition of LNA, DHA, EPA and n-3 PUFA of dorsal muscle and liver in VO-based diets increased with increasing LNA/LA ratio. In the liver and proximal intestine (PI), lysozyme (LZM) activity increased with the increasing LNA/LA ratio in the VO-based diet. However, it decreased in the mid-intestine (MI) and distal intestine (DI). Acid phosphatase (ACP) activity in liver was significantly high when the LNA/LA ratio in the diet was 1.06 (p < .05). Alkaline phosphatase (AKP) activity in the MI increased with the increasing dietary LNA/LA ratio. The LNA/LA ratio in the diet did not affect the growth of Manchurian trout juveniles, and a diet with an LNA/LA ratio between 0.55 and 1.06 was beneficial for non-specific immunity in Manchurian trout.

Динамика и современное состояние ихтиофауны реки Амазар после строительства гидроузла ЦПК «Полярная»

The ecosystem of the Amazar River, the first large tributary of the Upper Amur, has experienced considerable anthropogenous influence over the last 20 years. The construction of the water reservoir in the middle reaches of the river was the most extensive interference into the ecosystem. In 2018-2019 integrated hydrobiological, hydrochemical, and ichthyological research was conducted in the Amazar River during the navigation season. The article analyzes the current state of ichthyocenosis as the most vulnerable component of the ecosystem of the middle reaches of the river Amazar and the newly formed reservoir. The results of the ichthyological survey have established the presence of 15 species from 7 families. The new data are compared with the results of previous research. The authors make a conclusion concerning the changes which have occurred in the species composition in the Amazar River over a 27-year period. The initial data was collected on the ichthyofauna of the recently constructed water reservoir. It predominantly includes coarse fish (Lagowski’s minnow (Phoxinus lagowskii), Amur gudgeon (Gobio cynocephalus), Amur bitterling (Rhodeus sericeus), and Amur sleeper (Perccottus glenii). Manchurian trout (Brachymystax lenok) and Upper Amur grayling (Thymallus grubii) were observed in the reservoir only during spawning and downstream migration. The findings show that the ichthyofauna of the reservoir is formed by non-target fish. The largest diversity of species was observed in the downstream water of the Amazar River. It was determined that spawning migration through the fish pass extended from the first decade of May through the first decade of June. It was observed that at the beginning migration was dominated by older Manchurian trout and grayling, while at the end of the migration period there were mainly cyprinids and immature Manchurian trout and grayling. The authors suggest that the size of migrating fish should be taken into consideration to ensure the effective operation of the fish pass. Absence of water flow control in the fish pass and a lack of consideration for the size of fish may result in both, insufficient water level for large mature salmon fish, and prohibitively high speed of the current for the small cyprinids.

Ecological health assessments using fish communities in the habitat of Manchurian trout

Ecological health assessments using fish communities in the habitat of Manchurian trout

Influence of dietary linseed oil as substitution of fish oil on whole fish fatty acid composition, lipid metabolism and oxidative status of juvenile Manchurian trout, Brachymystax lenok

In this study, juvenile Manchurian trout, Brachymystax lenok (initial weight: 6.43 ± 0.02 g, mean ± SE) were received for nine weeks with five types of diets prepared by gradually replacing the proportion of fish oil (FO) with linseed oil (LO) from 0% (LO0) to 25% (LO25), 50% (LO50), 75% (LO75), and 100% (LO100). The eicosapentaenoic (EPA) and docosahexaenoic (DHA) composition decreased with increasing inclusion level of LO (P < 0.05). With increasing LO inclusion level, triglyceride (TAG) content of serum increased significantly, however, there was a decrease in high-density lipoprotein cholesterol (HDL) (P < 0.05). LO substitution of FO up-regulated the gene expression level of lipid metabolism-related genes Fatty Acid Desaturases 6 (FAD6), Acetyl-Coa Carboxylase (ACCα), Sterol Regulatory Element Binding Protein 1 (SREBP-1), and Sterol O- Acyl Transferase 2 (SOAT2), and down-regulated the gene expression level of Peroxisome Proliferator-Activated Receptor a (PPARα) (P < 0.05). The SOD activities of both serum and liver in LO100 were significantly lower than in LO25 (P < 0.05). The CAT activity of the liver in LO100 was significantly lower than in LO0 and LO25 (P < 0.05). This study indicates that the Manchurian trout may have the ability to synthesize LC-PUFAs from ALA, and an appropriate LO in substitution of FO (<75%) could improve both the lipid metabolism and the oxidation resistance.

Molecular Cloning, mRNA Expression, and Nutritional Regulation of a Fatty Acyl Δ6-Desaturase-like Gene of the Manchurian Trout, Brachymystax lenok (Pallas)

The fatty acyl delta-6 desaturase (Δ6-desaturase) is a key rate-limiting enzyme in the biosynthesis of long-chain polyunsaturated fatty acids (LCPUFAs). To study the effects of different oil sources in the feed on the nutritional regulation of Δ6-desaturase-like gene mRNA expression, the fulllength cDNA of the Δ6-desaturase-like gene was cloned from the liver of Manchurian trout (Brachymystax lenok). A 9-week feeding trial was performed, and the fish received diets with three different oil sources: fish oil (FO), sunflower oil (SO), and linseed oil (LO). The results showed that the 2448 bp long full-length cDNA contained an open reading frame (with a length of 1365 bp), encoding 454 amino acids. Gene expression analysis indicated that Δ6-desaturase-like gene mRNA is widely distributed throughout different tissues, with highest expression levels in both the liver and the brain. The linolenic acid (ALA) and linoleic acid (LA) compositions were highest in LO and SO diets, respectively. The eicosapentaenoic acid and docosahexaenoioc acid composition in LO and SO diets were significantly lower than in the FO diet. Fish fed with SO and LO showed significantly higher liver expression levels of Δ6-desaturase-like gene mRNA than those fed with FO. This indicates that SO and LO in the diet affected the nutritional regulation of the Δ6-desaturase-like gene mRNA in the liver of the Manchurian trout. In addition, these data suggest that the Manchurian trout has the ability to synthesize long-chain unsaturated fatty acids (LC-PUFAs) from ALA and LA.

Effect of variation in the dietary ratio of linseed oil to fish oil on growth, body composition, tissues fatty acid composition, flesh nutritional value and immune indices in Manchurian trout,Brachymystax lenok

In order to study the effects of linseed oil substitution on the growth, body composition, tissue fatty acid composition, flesh nutritional value and immune indices of juvenile Manchurian trout, five feed types containing different levels of linseed oil (LO) mixed with fish oil (FO) were prepared: 0 (0 LO); 250 g/kg (25 LO); 500 g/kg (50 LO); 750 g/kg (75 LO); and 1000 g/kg (100 LO); and fed to juvenile Manchurian trout (initial weight 6.43 ± 0.02 g) for 9 weeks. The results showed that substitution of FO with 750 g/kg LO did not affect the growth of juvenile trout, with protein content in the dorsal muscle, and lipid content in the liver not showing any significant difference (p > 0.05). The highest lipid content found in muscle samples occurred for the 25 LO diet. The fatty acid composition found in the dorsal muscle and the liver of the Manchurian trout reflects the fatty acid composition in the diet, where the relative amount of linolenic acid (ALA), linoleic acid (LA) and docosahexaenoic acid (DHA) found in these organs has a positive linear correlation with their relative composition in the diet (p < 0.05). As the amount of LO in the diet was increased, the composition of ALA found in the sampled organs increased, while the composition of DHA and eicosapentaenoic acid (EPA) decreased. At the same time, the index of atherogenicity (IA) and thrombogenicity (IT) of the muscle samples from the 75 LO and 100 LO diets was significantly lower than for the 0 LO and 25 LO diets (p < 0.05), while the flesh lipid quality (FLQ) in the 100 LO diet was significantly lower than for the other diets (p < 0.05). The aspartate transaminase (AST) activity decreased initially, and then increased, as the level of LO replacement for FO was increased, with the 25 LO diet being significantly lower than for other groups (p < 0.05). The alanine aminotransferase (ALT) activity in serum samples from the 100 LO diet was higher than that from other diets. The lysozyme (LZM) activity in both serum and liver tissue first increased to a peak for the 25 LO and 50 LO diets, respectively, and then decreased as the level of LO was further increased. There was no significant change in the alkaline phosphatase (AKP) activity in the liver samples; however, the acid phosphatase (ACP) activity decreased significantly from the highest value for 0 LO feed group. In conclusion, the composition of fatty acids in the dorsal muscle and the liver was found to be modified by the diets, and with the diet containing less than 750 g/kg LO, being both beneficial for growth, and improved immunity, while maintaining the nutritional value of the lipid content in the dorsal muscle during the 9‐week period.

Effects of dietary lipid levels on growth, feed utilization, digestive tract enzyme activity and lipid deposition of juvenile Manchurian trout,Brachymystax lenok(Pallas)

Six isoproteic diets were designated to evaluate the effects of dietary lipid levels (from 70 to 270 g/kg) on the growth performance, feed utilization, digestive tract enzyme activity and lipid deposition of juvenile Brachymystax lenok (average initial weight 0.54 ± 0.04 g). Each diet was fed to triplicate tanks (30 fish per tank) in an indoor closed recirculating system for 9 weeks. Final body weight and weight gain were highest in fish fed 190 g/kg diet and lowest in fish fed the 70 g/kg diet. Specific growth rate of fish fed with 190 g/kg diet was significantly higher than those fed with 70 and 270 g/kg diets (p < .05). Protein efficiency ratio of fish fed with 70 g/kg diet was significantly lower than the 110–230 g/kg treatments and was not significantly different from the 270 g/kg treatment. Fish fed with 270 g/kg diet had significantly higher hepatosomatic index and viscerosomatic index than those fed with 70–190 g/kg diets (p < .05). Intraperitoneal fat ratio and the whole-body lipid content had a trend to increase with increase in dietary lipid level. Muscle crude lipid content increased up to 190 g/kg with increase in dietary lipid level. Lipid retention decreased with increase in dietary lipid level, while no significant differences in protein intake and retention levels were observed in fish among all treatments. Lipase activity of the mixture of pyloric caeca and foregut in fish fed 190 and 230 g/kg diets was significantly higher than those fed 70 and 110 g/kg diets. Midgut and hindgut lipase activities of fish were significantly higher than those fed the 190 and 230 g/kg diets. In conclusion, based on the second-order polynomial model of WG and FCR, this study suggested that 173.8–195.0 g/kg dietary lipid levels were appropriated for B. lenok.

Genetic diversity and genetic structure of the endangered Manchurian trout, Brachymystax lenok tsinlingensis, at its southern range margin: conservation implications for future restoration

The Manchurian trout, Brachymystax lenok tsinlingensis (family: Salmonidae), is a cold freshwater fish endemic to Northeast Asia. South Korean populations of this species, which comprise its southern range limit, have recently decreased markedly in size and are now becoming critically endangered. We assessed the current population status of this species in South Korea by estimating the levels of genetic diversity and genetic structure of five natural and four restored populations using mitochondrial DNA (mtDNA) control region sequences and eight nuclear microsatellite loci. Levels of within-population genetic diversity were low, suggesting that past effective population sizes (N e) have been small. Each population had one or a maximum of two mtDNA haplotypes. Microsatellite allelic richness (AR) was significantly higher for natural populations (mean AR = 3.51; 95% confidence interval, 3.00–4.03) than for restored populations (mean AR = 2.61; 2.38–2.98). South Korean populations were significantly genetically isolated from one another, with private mtDNA haplotypes and microsatellite alleles, suggesting that limited gene flow has been occurring among populations. A mtDNA phylogeny revealed that South Korean lineages were more closely related to those of China than to those of North Korea and Russia. Overall, we suggest that future restoration efforts aimed at South Korean populations should consider the genetic characteristics reported here, which should help to fulfil effective conservation strategies for this highly cherished species. Our results will inform other conservation efforts, including assisted migration of freshwater fish populations at the equatorial end of the geographical range limit of the species.

Successful cryopreservation of spermatogonia in critically endangered Manchurian trout (Brachymystax lenok)

Because of the lack of cryopreservation techniques for fish eggs and embryos, cryopreservation of fish spermatogonia and subsequent generation of eggs and sperm would be an exit strategy for the long-term preservation of genetic resources. This study aimed to optimize cryoprotectants, cooling rates, and thawing temperatures for slow freezing of spermatogonia from endangered Manchurian trout (Brachymystax lenok). Whole testes were frozen with a cryomedium containing 1.3 M methanol, 0.2 M trehalose, and 10% egg yolk at a cooling rate of −1 °C/min and then stored in liquid nitrogen for 2 days. After thawing at 30 °C in a water bath, testicular cells from thawed testes were intraperitoneally transplanted into allogeneic triploid hatchlings. Transplanted spermatogonia migrated toward and were incorporated into recipient gonads, where they underwent gametogenesis. Transplantation efficiency did not significantly differ between frozen and fresh testes, demonstrating that Manchurian trout spermatogonia can be successfully cryopreserved in liquid nitrogen.

The complete mitochondrial genome of Brachymystax lenok tsinlingensis (Salmoninae, Salmonidae) and its intraspecific variation

The Manchurian trout, Brachymystax lenok tsinlingensis, is endangered in Korea, where the southern range limit for this cold-freshwater fish occurs. In this study, the complete mitochondrial genome of Korean B. lenok tsinlingensis was sequenced and its genetic characteristics were identified. The mitogenome of B. lenok tsinlingensis comprises 16,748 base pairs containing 37 genes (13 protein-coding genes, 22 tRNA genes, and 2 rRNA genes) and one major non-coding region (control region), making it similar to the majority of vertebrate mitogenomes. Interestingly, at the base of the stem region of OL in B. lenok tsinlingensis, the conserved motif is replaced by a 5′-ACCGG-3′ motif instead of the 5′-GCCGG-3′. We also identified an 81-base-pair tandem-repeat motif in the control region, the length of which is reduced by one nucleotide compared to those in B. lenok and Hucho species. The number of repeat motifs differed between Korean and Chinese B. lenok tsinlingensis, with two and three reiterations, respectively. The control region of B. lenok and its relatives will be used as a genetic marker in evolution/genetic studies and as a PCR-based marker for rapid identification of their lineages.

Growth, feed utilization and body composition of juvenile Manchurian trout,Brachymystax lenok(Pallas) fed different dietary protein and lipid levels

A 10-week feeding trial with four dietary protein levels (400, 450, 500 and 550 g kg−1 crude protein) and two dietary lipid levels (80 and 160 g kg−1 crude lipid) was conducted to assess optimum dietary protein and lipid levels for the growth, feed utilization and body composition of juvenile Manchurian trout (initial weight 11.80 ± 0.15 g). Fish were fed twice daily (08:30 and 16:30 h) to apparent satiation. The results showed that fish fed the diet with 500 g kg−1 protein and 80 g kg−1 lipid had the highest growth and feed efficiency. However, fish fed the diet with 450 g kg−1 protein and 160 g kg−1 lipid showed comparable growth to that of the fish fed diet 5 (500/80) and had higher protein efficiency ratio (PER), nitrogen retention (NR) and energy retention (ER) than other groups (P < 0.05). Growth, PER, NR and ER of fish fed the 160 g kg−1 lipid diet was significantly higher (P < 0.05) than that of fish fed the 80 g kg−1 lipid diet at 400 and 450 g kg−1 protein diet, whereas these values showed an opposite trend at 500 and 550 g kg−1 protein diet, and the lowest PER, NR and ER was found by fish fed the 400 g kg−1 protein diet with 80 g kg−1 lipid. Fish fed diets with 400 g kg−1 protein had lower feed intake (FI) than that of other groups. Feed intake of fish fed 80 g kg−1 lipid level was significantly lower than that of fish fed 160 g kg−1 lipid diet at 400 g kg−1 protein (P < 0.05), while no significant differences were observed at 450, 500 and 550 g kg−1 protein-based diets. Contrary to moisture content, lipid content of whole body and muscle increased significantly (P < 0.05) with increasing lipid levels. The results of this study indicated that the diet containing 450 g kg−1 protein and 160 g kg−1 lipid, with a P/E ratio of 23.68 g protein MJ−1 would be suitable for better growth and feed utilization of juvenile Manchurian trout under the experimental conditions and design level used in this study.

Production and characterization of recombinant Manchurian trout thyrotropin

Thyrotropin (thyroid-stimulating hormone, TSH), a heterodimeric glycoprotein hormone produced in the pituitary, stimulates the thyroid gland and release of thyroid hormones. In contrast to a well-known efficacy of recombinant mammalian TSHs, there is no report about the production of teleost recombinant TSH and its biological activity. In this study, we report the production of a single-chain recombinant TSH (mtTSH) of Manchurian trout (Brachymystax lenok), by baculovirus in silkworm (Bombyx mori) larvae. The mtTSH was produced in silkworm larvae and characterized as a form of N-linked glycosylation. The cAMP signaling system in transiently transfected COS-7 cells revealed that the mtTSH was recognized by their cognate receptors, salmon TSHα and TSHβ receptors, but not LH receptor. The thyrotropic potency of the mtTSH was examined by rainbow trout basibranchial tissues containing thyroid follicles. The height of follicle epithelial cells was significantly increased by treatments of mtTSH in vivo and in vitro. In conclusion, the present study suggests that the mtTSH produced by baculovirus-silkworm larvae is a biologically active recombinant TSH.

Steroidogenic response of carp ovaries to piscine FSH and LH depends on the reproductive phase

The gonadotropins (GTHs) follicle-stimulating hormone (FSH) and luteinizing hormone (LH) are the key regulators of reproduction. We determined the competence of heterologous recombinant GTHs at eliciting steroid secretion from carp ovaries at different reproductive stages. We collected carp ovaries at: early, mid and end vitellogenesis, when most of the oocytes still contained a germinal vesicle (GV) at a central stage, and mature ovaries with a migrating GV. Plasma estradiol (E2) levels at early vitellogenesis were high and decreased thereafter. Basal secretion levels of E2 increased with oocyte diameter and GSI value, whereas 17α,20β-dihydroxy-4-pregnen-3-one (DHP) was detected only in females with mature follicles.Carp ovary fragments were exposed to recombinant fish GTHs belonging to different teleost orders: Japanese eel (Anguilla japonica, Anguilliformes), Manchurian trout (Brachymystax lenok, Salmoniformes), and Nile tilapia (Oreochromis niloticus); to mammalian GTHs (pFSH and hCG), or to carp and tilapia pituitary extract (CPE and TPE, respectively).All of the recombinant GTHs tested stimulated steroid secretion. However, the steroid secretion differed according to the type of GTH and the developmental state of the ovary. CPE increased the secretion of both E2 and DHP at almost all stages of ovarian maturity. In mature ovarian fragments, DHP secretion was higher in response to recombinant LHs (eel and tilapia) than to recombinant FSH. Early- and mid-vitellogenic ovaries showed no secretion of DHP and high secretion of E2 in response to all recombinant GTHs tested. This is in line with the hypothesis that LH regulates the final stages of maturation, when the involvement of FSH is marginal. These results may contribute to understanding the mechanisms that determine differential activation of steroid secretion and specificity in fish.

Perspectives on fish gonadotropins and their receptors

Teleosts lack a hypophyseal portal system and hence neurohormones are carried by nerve fibers from the preoptic region to the pituitary. The various cell types in the teleost pituitary are organized in discrete domains. Fish possess two gonadotropins (GtH) similar to FSH and LH in other vertebrates; they are heterodimeric hormones that consist of a common α subunit non-covalently associated with a hormone-specific β subunit. In recent years the availability of molecular cloning techniques allowed the isolation of the genes coding for the GtH subunits in 56 fish species representing at least 14 teleost orders. Advanced molecular engineering provides the technology to produce recombinant GtHs from isolated cDNAs. Various expression systems have been used for the production of recombinant proteins. Recombinant fish GtHs were produced for carp, seabream, channel and African catfish, goldfish, eel, tilapia, zebrafish, Manchurian trout and Orange-spotted grouper. The hypothalamus in fishes exerts its regulation on the release of the GtHs via several neurohormones such as GnRH, dopamine, GABA, PACAP, IGF-I, norepinephrine, NPY, kisspeptin, leptin and ghrelin. In addition, gonadal steroids and peptides exert their effects on the gonadotropins either directly or via the hypothalamus. All these are discussed in detail in this review. In mammals, the biological activities of FSH and LH are directed to different gonadal target cells through the cell-specific expression of the FSH receptor (FSHR) and LH receptor (LHR), respectively, and the interaction between each gonadotropin-receptor couple is highly selective. In contrast, the bioactivity of fish gonadotropins seems to be less specific as a result of promiscuous hormone–receptor interactions, while FSHR expression in Leydig cells explains the strong steroidogenic activity of FSH in certain fish species.

Dietary Lipid Level Induced Antioxidant Response in Manchurian Trout, Brachymystax lenok (Pallas) Larvae

This study was designed to determine the nutritional lipid requirement of Manchurian trout and to investigate the effects of lipid concentrations on the antioxidant status in larvae with experimental diets with different lipid levels. Oxidative stress differences between different organs and tissues were also assessed. Manchurian trout larvae were fed for 35 days and, during that period, growth and survival, the activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and content of malondialdehyde (MDA) in viscera, muscle, gill and brain of four diets, lipid levels from 15 to 30%, and control treatment were measured. Growth rates were similar, but survival was low, between high and low dietary lipid levels. SOD activity was stimulated in viscera, muscle and brain in high lipid diets, but reduced in gills with increased lipid content. SOD was kept lower in the control group. GPX activity was inhibited in viscera and stimulated in gill, muscle and brain. CAT activities were enhanced by all treatments and showed the lowest values in the control. Lipid peroxidation of the diet was promoted in all organs, excluding the gill which showed no regular pattern. MDA content increased with increased dietary lipid levels in viscera, muscle and brain. Our results indicate that the most appropriate lipid requirement is probably 20-25% and a higher dietary level of lipids might induce oxidative stress in Manchurian trout larvae. The brain and gill were probably the most sensitive organs to oxidative damage.

Biological activities of recombinant Manchurian trout FSH and LH: their receptor specificity, steroidogenic and vitellogenic potencies

Gonadotropins (GTHs), FSH and LH, play central roles in vertebrate reproduction. Here, we report the production of biologically-active recombinant FSH (r-mtFSH) and LH (r-mtLH) of an endangered salmon species, Manchurian trout (Brachymystax lenok), by baculovirus in silkworm (Bombyx mori) larvae. The biological activities of the recombinant hormones were analyzed using COS-7 cell line transiently expressing either amago salmon FSH or LH receptor. The steroidogenic potency of the r-mtFSH and r-mtLH was examined by a culture system using rainbow trout follicles in vitro. In vivo, bioactivity was assessed by measuring ovarian weight, oocyte diameter, and plasma steroid hormone levels in female rainbow trout. Moreover, inducing potency of milt production were examined in vivo using goldfish. Our results demonstrated that the r-mtFSH and r-mtLH were successfully produced in the baculovirus-silkworm system and recognized by their cognate receptors specifically in vitro. The production of estradiol-17beta (E2) and testosterone (T) was stimulated by the r-mtFSH and r-mtLH respectively, from the full-grown follicles of rainbow trout, whereas both E2 and T were increased by relatively higher doses of the recombinant hormones from the follicles of the maturing stage. In in vivo assay, injection of the r-mtFSH but not r-mtLH increased ovarian weight, oocyte diameter, and plasma E2 levels in immature rainbow trout. Injection of both r-mtFSH and r-mtLH induced milt production in male goldfish. In conclusion, the present study strongly suggests that the r-mtFSH and r-mtLH have distinct biological properties, such as a specific responsiveness for the cognate receptor, steroidogenic, and vitellogenic activities for ovarian follicles in salmonids. These recombinant FSH and LH may be applied for future studies on the gonadal development and maturation in fishes as well as the endangered salmon species.

18S rRNA gene sequences and random amplified polymorphic DNA used in discriminating Manchurian trout from other freshwater salmonids

18S rRNA gene sequences and random amplified polymorphic DNA used in discriminating Manchurian trout from other freshwater salmonids