Exploring the effects of dietary inulin in rainbow trout fed a high‑starch,100%plant‑based diet

Background High dietary carbohydrates can spare protein in rainbow trout(Oncorhynchus mykiss)but may affect growth and health.Inulin,a prebiotic,could have nutritional and metabolic effects,along with anti-inflammatory properties in teleosts,improving growth and welfare.We tested this hypothesis in rainbow trout by feeding them a 100%plant-based diet,which is a viable alternative to fishmeal and fish oil in aquaculture feeds.In a two-factor design,we examined the impact of inulin(2%)as well as the variation in the carbohydrates(CHO)/plant protein ratio on rainbow trout.We assessed the influence of these factors on zootechnical parameters,plasma metabolites,gut microbiota,production of short-chain fatty acids and lactic acid,as well as the expression of free-fatty acid receptor genes in the mid-intestine,intermediary liver metabolism,and immune markers in a 12-week feeding trial.Results The use of 2%inulin did not significantly change the fish intestinal microbiota,but interestingly,the high CHO/protein ratio group showed a change in intestinal microbiota and in particular the beta diversity,with 21 bacterial genera affected,including Ralstonia,Bacillus,and 11 lactic-acid producing bacteria.There were higher levels of butyric,and valeric acid in groups fed with high CHO/protein diet but not with inulin.The high CHO/protein group showed a decrease in the expression of pro-inflammatory cytokines(il1b,il8,and tnfa)in liver and a lower expression of the genes coding for tight-junction proteins in mid-intestine(tjp1a and tjp3).However,the 2%inulin did not modify the expression of plasma immune markers.Finally,inulin induced a negative effect on rainbow trout growth performance irrespective of the dietary carbohydrates.Conclusions With a 100%plant-based diet,inclusion of high levels of carbohydrates could be a promising way for fish nutrition in aquaculture through a protein sparing effect whereas the supplementation of 2%inulin does not appear to improve the use of CHO when combined with a 100%plant-based diet.

Short-Term Stress Response of Juvenile Rainbow Trout Subjected to Two Different Rearing Densities

Juvenile rainbow trout Oncorhynchus mykiss were subjected to one of four treatments in a two-by-two experimental design: 1) fed at a density of 1.8 g/m3, 2) Fasted at 1.8 g/m3, 3) fed at 30.1 g/m3, and 4) fasted at 30.1 g/m3. Blood glucose and hematocrit were measured at 4, 6, 48, 168, and 336 hours after placement in one of the two rearing densities, with relative fin lengths and organosomatic indices recorded at 336 hours. Glucose levels over time were not significantly different among the density and feeding treatments. Hematocrit levels over time were also not significantly different. Total lengths, weight, hepatosomatic index, viscerosomatic index, and any of the relative fin lengths were not significantly different between the high and low densities. However, the hepatosomatic index was significantly greater in the fed fish compared to those fasted. The splenosomatic index was significantly greater in the higher density treatment. These results likely indicate no short-term stress response to the higher rearing density used in this short-term experiment and no interaction between starvation and density-related stressors.

IMMUNE FUNCTION OF IMMATURE RAINBOW TROUT ( Oncorhynchus mykiss ) RELATED WITH PHOTOPERIOD

Radioimmunoassay,enzyme linked immunospot assay and enzyme immunoassay were used for the determinations of plasma steroid hormone's level,antibody producing cell's counting and IgM level in this study.The decreased number of antibody producing cells and low IgM levels were observed in sexual immature rainbow trout during the spawning season.These fish were reared under almost constant water temperature and natural photoperiod.Moreover,low IgM level was also observed in immature rainbow trout,which were reared under short photoperiod,and IgM level was not changed by treatment of testosterone.The results suggest that photoperoid may cause the changes in immune competence.It is possible that circadian rhythm accompanied with photoperiod may influence physiological function of fish,so that immune competence is changed.

17α,20β-DIHYDROXY-4-PREGNENE-3-ONE SUPPRESSES PLASMA IgM LEVELS IN RAINBOW TROUT (Oncorhynchus mykiss)

The relationships between 17α,20β dihydroxy 4 pregnene 3 one (17α,20β DP) and plasma IgM and total protein levels were investigated in rainbow trout, Oncorhynchus mykiss .IgM and total protein levels decreased in both sexes of mature rainbow trout when 17α,20β DP levels increased during the spawning season,while the elevated 17α,20β DP suppressed IgM levels and reversibly enhanced total protein concentrations in immature trout.This represents the first report of the effect of 17α,20β DP on fish immunity.These data suggest that the increase of 17α,20β DP in spawning season may be related to infectious skin diseases.

Screening of Saprolegniasis Preventive Drugs for Golden Trout (Oncorhynchus mykiss) Fertilized Eggs During Hatching

In order to screen the effective preventive drugs for saprolegniasis occurred during hatching of the fertilized eggs of golden trout, effects of two kinds of self-made Chinese herbal compounds（AB） and two kinds of chemical bactericides（peroxyacetic acid andhydrogen peroxide, CD） with three disinfection concentrations were compared in this study. The fertilized eggs of golden trout were randomly divided into 13 groups, each group with two parallels of 1 000 eggs. Drugs at high,medium and low concentration were respectively used to immerse the fertilized eggs35 min for disinfection once a day. The numbers of live and dead eggs were recorded each day before disinfection and the numbers of survival eyed eggs were counted in each experimental group after finishing the whole test. The results indicated that all of four kinds of drugs used here had a certain effect on prevention and control of saprolegniasis during hatching of fertilized eggs of golden trout, and also showed a correlation between the concentrations and effects. After the selection of several drugs, we found that group B-medium, C-medium and D-high exhibited the most remarkable effects on control of saprolegniasis. However, careful consideration should be given to chemical drugs residues and their potential hazard to the eyed eggs. Therefore, Chinese herbal compound B was suggested to be used for prevention and control of saprolegniasis to achieve the healthy aquaculture. And the concentration and operation time of Chinese herbal compound B should be strictly controlled when used in disinfection of fertilized eggs.

Thermal Tempering Does Not Impact Rainbow Trout and Brown Trout Survival

Hatchery-reared fish are frequently adjusted (tempered) to the higher water temperatures present in the water bodies where they are to be stocked. This study was undertaken to determine the necessity of such tempering practices. This study used rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta) reared at 11.2°C. The first two trials for each species were designed to simultaneously determine the upper incipient lethal temperature and 100% lethal temperature over a 14-day period. The third trial for each species evaluated the effects of an exaggerated 12-hour tempering regime on fish survival after placement in elevated water temperatures. After transfer from a water temperature of 11.2°C, no rainbow trout survived at 26°C, and only 50% survived at 25°C. No brown trout survived at 22°C and only 50% at 20°C. Survival of rainbow trout was not improved by the 12-hour tempering regime where water temperatures were slowly increased from 11.2°C to either 25°C or 26°C. Similarly, tempering did not improve brown trout survival at either 20°C or 22°C. These results suggest that tempering is not needed when hatchery-reared trout are reared and stocked at the water temperatures within the range of those used in this study.

Immune response,stress resistance and bacterial challenge in juvenile rainbow trouts Oncorhynchus mykiss fed diets containing chitosan-oligosaccharides

Effects of dietary supplementation of chitosan-oligosaccharides （COS） on the growth performance, immune response, stress resistance, and disease resistance of juvenile rainbow trout Oncorhynchus mykiss were studied. Four experimental diets containing 0, 20, 40, or 60 mg/kg COS （COSO, COS20, COS40, and COS60, respectively） were fed to juvenile rainbow trout （initial weight = 5.2 ± 0.3 g） for 8 weeks. By the end of the feeding trial, representative groups of fish from each dietary treatment were challenged with stressor （30 see air exposure） and pathogen exposure （intraperitoneal injection with Aeromonas hydrophila ）. Results showed that supplementation of COS in diets did not affect production performance and body composition of rainbow trout. However, fish fed the COS40 diet demonstrated improved phagocytic activities, respiratory burst activities and decreased serum cortisol level. Additionally, survival following A. hydrophila challenge was significant higher among fish fed the COS-supplemented feeds, although there was no difference based on the level of supplementation. The present study suggests that COS can be used as an immuno-stimulant in rainbow trout feeds

An initial investigation replacing fish meal with a commercial fermented soybean meal product in the diets of juvenile rainbow trout

The inclusion of PepSoyGen (PSG), a commercially-available fermented soybean meal product, was evaluated with juvenile rainbow trout, Oncorhynchus mykiss in an initial 70-day feeding trial, with a supplemental trial involving a subset of the experimental diets continuing for an additional 40 d. Six diets containing 0%, 10%, 20%, 30%, 40%, or 50% PSG, with the PSG directly replacing fish meal, were used in the first trial. There were no significant differences in weight gain or feed conversion ratio between the fish meal-based control diet and diets containing up to 30% PSG. However, weight gain was significantly reduced and feed conversion ratio significantly increased with the 40% and 50% PSG diets. No health assessment differences were observed in fish receiving any of the diets, and no evidence of gross gut inflammation was evident. There were no significant differences in weight gain or feed conversion ratio among the four dietary treatments ranging from 0% to 30% PSG which were fed for an additional 40 d after the initial 70-d trial (110 days total). Based on these results, juvenile rainbow trout diets can contain up to 30% PSG without any loss of rearing performance, thereby replacing at least 60% of the fish meal.

Effects of full replacement of dietary fishmeal with insect meal from Tenebrio molitor on rainbow trout gut and skin microbiota

Background:Aquaculture must continue to reduce dependence on fishmeal(FM)and fishoil in feeds to ensure sustainable sector growth.Therefore,the use of novel aquaculture feed ingredients is growing.In this regard,insects can represent a new world of sustainable and protein-rich ingredients for farmed fish feeds.Accordingly,we investigated the effects of full replacement of FM with Tenebrio molitor(TM)larvae meal in the diet of rainbow trout(Oncorhynchus mykiss)on fish gut and skin microbiota.Methods:A feeding trial was conducted with 126 trout of about 80 g mean initial weight that were fed for 22 weeks with two isonitrogenous,isolipidic,and isoenergetic extruded experimental diets.Partially defatted TM meal was included in one of the diets to replace 100%(TM 100)of FM,whereas the other diet(TM 0)was without TM.To analyse the microbial communities,the Illumina MiSeq platform for sequencing of 16S rRNA gene and Qiime pipeline were used to identify bacteria in the gut and skin mucosa,and in the diets.Results:The data showed no major effects of full FM substitution with TM meal on bacterial species richness and diversity in both,gut mucosa-and skin mucus-associated microbiome.Skin microbiome was dominated by phylum Proteobacteria and especially by Gammaproteobacteria class that constituted approximately half of the bacterial taxa found.The two dietary fish groups did not display distinctive features,except for a decrease in the relative abundance of Deefgea genus(family Neisseriaceae)in trout fed with insect meal.The metagenomic analysis of the gut mucosa indicated that Tenericutes was the most abundant phylum,regardless of the diet.Specifically,within this phylum,the Mollicutes,mainly represented by Mycoplasmataceae family,were the dominant class.However,we observed only a weak dietary modulation of intestinal bacterial communities.The only changes due to full FM replacement with TM meal were a decreased number of Proteobacteria and a reduced number of taxa assigned to Ruminococcaceae and Neisseriaceae families.Conclusions:The data demonstrated that TM larvae meal is a valid alternative animal protein to replace FM in the aquafeeds.Only slight gut and skin microbiota changes occurred in rainbow trout after total FM replacement with insect meal.The mapping of the trout skin microbiota represents a novel contribution of the present study.Indeed,in contrast to the increasing knowledge on gut microbiota,the skin microbiota of major farmed fish species remains largely unmapped but it deserves thorough consideration.

GFAP expression in the optic nerve and increased Н2S generation in the integration centers of the rainbow trout（Oncorhynchus mykiss） brain after unilateral eye injury

Hydrogen sulfide(H2S)is considered as a protective factor against cardiovascular disorders.However,there are few reports on the effects of H2S in the central nervous system during stress or injury.Previous studies on goldfish have shown that astrocytic response occurs in the damaged and contralateral optic nerves.Glial fibrillary acidic protein(GFAP)concentration in the optic nerves of rainbow trout has not been measured previously.This study further characterized the astrocytic response in the optic nerve and the brain of a rainbow trout(Oncorhynchus mykiss)after unilateral eye injury and estimated the amount of H2S-producing enzyme cystathionineβ-synthase(CBS)in the brain of the rainbow trout.Within 1 week after unilateral eye injury,a protein band corresponding to a molecular weight of 50 kDa was identified in the ipsi-and contralateral optic nerves of the rainbow trout.The concentration of GFAP in the injured optic nerve increased compared to the protein concentration on the contralateral side.The results of a quantitative analysis of GFAP+cell distribution in the contralateral optic nerve showed the largest number of GFAP+cells and fibers in the optic nerve head.In the damaged optic nerve,patterns of GFAP+cell migration and large GFAP+bipolar activated astrocytes were detected at 1 week after unilateral eye injury.The study of H2S-producing system after unilateral eye injury in the rainbow trout was conducted using enzyme-linked immunosorbent assay,western blot analysis,and immunohistochemistry of polyclonal antibodies again st CBS in the integrative centers of the brain:telencephalon,optic tectum,and cerebellum.Enzyme-linked immunosorbent assay results showed a 1.7-fold increase in CBS expression in the rainbow trout brain at 1 week after unilateral eye injury compared with that in intact animals.In the ventricular and subventricular regions of the rainbow trout telencephalon,CBS+radial glia and neuroepithelial cells were identified.After unilateral eye injury,the number of CBS+neuroepithelial cells in the pallial and subpallial periventricular regions of the telencephalon increased.In the optic tectum,unilateral eye injury led to an increase in CBS expression in radial glial cells;simultaneously,the number of CBS+neuroepithelial cells decreased in intact animals.In the cerebellum of the rainbow trout,neuroglial interrelationships were revealed,where H2S was released,apparently,from astrocyte-like cells.The organization of H2S-producing cell complexes suggests that,the amount of glutamate produced in the rainbow trout cerebellum and its reuptake was controlled by astrocyte-like cells,reducing its excitotoxicity.In the dorsal matrix zone and granular eminences of the rainbow trout cerebellum,CBS was expressed in neuroepithelial cells.After unilateral eye injury,the level of CBS activity increased in all parts of the cerebellum.An increase in the number of H2S-producing cells was a response to oxidative stress after unilateral eye injury,and the overproduction of H2S in the cerebellum occurred to neutralize reactive oxygen species,providing the cells of the rainbow trout cerebellum with a protective effect.A structural reorganization in the dorsal matrix zone,associated with the appearance of an additional CBS+apical zone,and a decrease in the enzyme activity in the dorsal matrix zone,was revealed in the zones of constitutive neurogenesis.All experiments were approved by the Commission on Biomedical Ethics,A.V.Zhirmunsky National Scientific Center of Marine Biology(NSCMB),Far Eastern Branch,Russian Academy of Science(FEB RAS)(approval No.1)on July 31,2019.

Comparisons of Salinity Adaptation in Terms of Growth, Body Composition, and Energy Budget in Juveniles of Rainbow and Steelhead Trouts (Oncorhynchus mykiss)

This study examined the effect of different salinities(0, 5, 10, 15, 20, 25 and 30) on the growth performance and energy budget of juveniles of two different ecotypes of Oncorhynchus mykiss, landlocked rainbow trout and anadromous steelhead trout. In the 42 d experiment, fish were cultured in three replicate tanks per salinity treatment(eight fish per tank). At the end of the experiment, the growth of rainbow and steelhead trouts was significantly higher at salinities of 5 and 10, respectively, than at all other salinities. The protein, lipid and energy content of both ecotypes declined with the increase of salinity. Based on their energy budgets, the percentage of energy consumed for growth by rainbow and steelhead trouts were significantly higher at salinities of 5(34.00% ±1.69%) and 10(43.76% ± 1.29%), respectively, than at all other salinities. The percentage of energy consumed for respiration by rainbow and steelhead trouts was lower at salinities of 5(54.90% ± 1.77%) and 10(46.73% ± 0.62%), respectively, than at all other salinities. Our results indicated that the salinity adaptation ability of juvenile steelhead trout was slightly better than that of juvenile rainbow trout, and salinities of 10 and 5, respectively, were most suitable for growth of these two fishes.

Temperature-Dependent Fatty Acid Composition Change of Phospholipid in Steelhead Trout (Oncorhynchus mykiss) Tissues

In this study, the changes of the fatty acid composition of phospholipid in different tissues(muscle, heart, brain and spleen) of steelhead trout(Oncorhynchus mykiss) were analyzed when the water temperature decreased gradually from 16 to 12℃℃, 8℃, 6℃, 4℃, 2 and 1℃℃. Three fish individuals each tank(average weight 70.32 g ± 9.12 g) were collected and used to analysis at each designed temperatures. At normal temperature(16℃), the fatty acid composition of phospholipid of muscle and heart was similar each other. The highest concentration of saturate fatty acids(SFA) was found in the phospholipid of spleen. The brain phospholipid contained higher oleic acid(18:1 n9) than the phospholipid of other tissues at 16℃. When the environmental temperature decreased, the concentration of unsaturated fatty acids of phospholipids in all tissues increased, and accordingly the ratio pf the unsaturated to saturated fatty acids(U/S) and unsaturation index(UI) increased, indicating that steelhead trout can compensate temperature-dependent changes in membrane fluidity by remodeling the fatty acid composition of phospholipids. The changes in the fatty acid composition of phospholipid were tissue-specific. At the early stages of the experiment(16 to 8℃℃), the fatty acid composition of phospholipid changed remarkably in muscle, heart, and spleen. When temperature decreased to less than 8℃, an obvious response of phospholipid fatty acid was observed in all tissues. The change of phospholipid composition of steelhead trout tissues may be affected by both cold stress and starvation when the temperature decreased to 2℃, and the change of phospholipid composition of muscle was very obvious.

An Effective Method of Prompting Juvenile Rainbow Trout (Oncorhynchus mykiss) to Cope with Heat Stress

In the present study,juvenile rainbow trout(Oncorhynchus mykiss)were acclimated at 5 temperatures.Fish in the control group(C0)were reared at a constant temperature(16℃);trout in four other treatments(A0,A3,A6,and A9)were acclimated to a high temperature(22℃)for 0,3,6,and 9 d,respectively,and then returned to normal temperature(16℃)for 7d.The temperature was then raised to 20℃and the fish were cultured for 40 d.The results showed that the growth rates of A3,A6,and A9 were higher than that of A0 but lower than that of C0.The growth rate of A9 was the highest among the 4 acclimation groups.The activities of serum transaminase and liver antioxidant enzyme significantly increased during acclimation.At the beginning of growth,the activities of enzymes were lower in A3,A6,and A9 with the lowest in A9.In addition,the transcript abundance of heat shock protein(HSP)60 gene in A9 was not significantly different from that of C0 during growth.HSP70 in A9 significantly increased at the beginning and returned to that of C0 at the end.Our findings indicated that pre-acclimation improved the high temperature tolerance with the best effectiveness observed at 22℃for 9 d.A possible mechanism underlining such phenomenon is the improvement of antioxidant defense system.

Effect of micro-algae Schizochytrium sp.supplementation in plant diet on reproduction of female rainbow trout(Oncorhynchus mykiss):maternal programming impact of progeny

Background:The broodstock diet,and in particular the lipid and fatty acid composition of the diet,is known to play a key role in reproductive efficiency and survival of the progeny in fish.A major problem when replacing both fish meal and fish oil by plant sources is the lack of n-3 long chain polyunsaturated fatty acids,such as eicosapentaenoic acid(EPA)and docosahexaenoic acid(DHA).To address this problem,we studied the effect of the plant-based diet supplemented with Schizochytrium sp.microalgae,source of DHA,compared to a conventional commercial diet rich in fish meal and fish oil on reproductive performance and egg quality and the consequences on progeny,in female rainbow trout broodstock.Results:The results demonstrated that DHA-rich microalgae supplementation in a plant-based diet allowed for the maintenance of reproductive performance and egg quality comparable to a conventional commercial feed rich in fish meal and fish oil and led to an increased significant fry survival after resorption.Moreover,when females were fed a plant-based diet supplemented with micro-algae,the 4-month-old progenies showed a significant higher growth when they were challenged with a similar diet as broodstock during 1 month.We provide evidence for metabolic programming in which the maternal dietary induced significant protracted effects on lipid metabolism of progeny.Conclusions:The present study demonstrates that supplementation of a plant-based diet with DHA-rich microalgae can be an effective alternative to fish meal and fish oil in rainbow trout broodstock aquafeed.

Influence of Maternal and Larval Immunisation against <i>Lactococcus garviae</i>Infection in Rainbow Trout <i>Oncorhynchus mykiss</i>(Walaum) Lysozyme Activity and IgM Level

This study evaluated efficacy of maternal and larval immunisation against Lactococcus garviae infection and on the lysozyme and immunoglobulin (IgM) levels in rainbow trout Oncorhynchus mykiss (Walaum). Forty-eight-day-old larvae (mean weight 96 mg) originating from injected weekly with letrozole and immunised, only immunised and non-immunised parents were experimentally infected with the L. garvieae, and the mortality rate was recorded daily. Larvae were vaccinated by immersion at 58 days post hatch with live L. garvieae (109 cells/mL) for 15 min. Every third day post larvae vaccination, two larvae from each group were collected for analysis lysozyme (by a method based on the ability of lysozyme to lyse the bacterium Micrococcus lysodeikticus) and IgM (by enzyme-linked immunosorbent assay (ELISA)) parameters. Vaccinated and control larvae were tested for protection against L. garvieae 30 days post larvae immunization when the larvae were 88 days old. Larvae were challenged by bath exposure with live L. garvieae (109 cells/mL) for 2 min and monitored for mortality for at least 10 days following challenge. The challenge experiment with L. garvieae showed a significant reduction in larvae from immunised (54.44% ± 0.64%) and injected weekly with letrozole and immunised fish (52.96% ± 0.97%) compared to larvae from control fish (62.96% ± 2.22%). Vaccinated larvae originated from injected weekly with letrozole and immunised parents showed significantly higher lysozyme activity compared to other fish groups. Vaccinated larvae showed significantly less mortality compared to controls. The relative percent survival (RPS) values of larvae from only immunised, injected weekly with letrozole and immunised and non-immunised parents vaccinated with L. garvieae were 67.36% ± 0.9%, 68.05% ± 0.66% and 48.27% ± 2.79% respectively. The results indicate that the effect of maternal immunization rainbow trout against L. garvieae infection by eliciting the immune responses as indicated by an increase in the IgM level and lysozyme activity.

Rearing Performance of Juvenile Brown Trout (<i>Salmo trutta</i>) Subjected to Exercise and Dietary Bioprocessed Soybean Meal

This 121-day experiment evaluated the rearing performance of brown trout Salmo trutta fed one of two isonitrogenous and isocaloric diets and reared at velocities of either 2.8 or 16.1 cm/s. Fishmeal was the primary protein source for the reference diet, and bioprocessed soybean meal replaced approximately 67% of the fishmeal in the experimental diet. At the end of the experiment, there were no significant differences in gain, percent gain, feed conversion rates, or specific growth rates between the dietary treatments. There were also no significant differences in intestinal morphology, splenosomatic, hepatosomatic, and viscerosomatic indices related to diet composition. However, gain, percent gain, feed fed, and specific growth rate were all significantly greater in brown trout reared at the higher velocity. No significant differences in any of the other variables measured were observed between the velocity treatments. There were no significant interactions between diet and velocity in any of the variables. Based on the results of this study, bioprocessed soybean meal can replace at least 67% of the fishmeal in brown trout diets, regardless of the rearing velocities used in this study. However, higher rearing velocities are recommended to maximize juvenile brown trout growth rates.

Brown Trout Spawn Timing, Redd Locations, and Stream Characteristics in Spearfish Creek within Spearfish, South Dakota, USA

This study documented brown trout (Salmo trutta) spawning locations, redd construction timing, and associated environmental variables in an 850-m long mainstem section and a 400-m long diversion channel of Spearfish Creek within the city limits of Spearfish, South Dakota, USA in 2019. The first redds were observed on October 15, with no new redds observed after November 12. Redd construction peaked during the first week of November, when 23 redds were observed in the mainstem section and 50 in the diversion channel. Substrate size was significantly smaller, water temperatures significantly higher, and water velocities significantly greater in redd versus non-redd locations in both the mainstem reach and the diversion channel (P &#730;C), but significant, increase in water temperature from the rest of the channel locations. This is the first study to document redd locations in Spearfish Creek and will provide a baseline to evaluate future spawning activity, particularly as it may be affected by likely future anthropogenic changes potentially affecting the stream environment.

ANNUAL CHANGES IN PLASMA LEVELS OF CORTISOL AND SEX STEROID HORMONES IN MALE RAINBOW TROUT, ONCORHYNCHUS MYKISS

The profiles of cortisol, testosterone, 11 ketotestosterone and 17α, 20β dihydroxy 4 pregnene 3 one in male rainbow trout reared under constant water temperature and natural photoperiod were determined by radioimmunoassay. Gonads of male rainbow trout reached maturity when the fish were two years old. Changes in the plasma levels of both sex steroid hormones and cortisol were closely related to the GSI. Plasma levels of testosterone, 11 ketotestosterone and 17α, 20β dihydroxy 4 pregnene 3 one showed a clear peak in the annual breeding season, when the GSI reached their maxima. Plasma cortisol levels also showed clearly seasonal changes in both two and three year old fish. The results suggest that the elevated plasma levels of cortisol may not just be due to stresses during the breeding season but have certain physiological functions in the reproduction of rainbow trout.

Trout Redd Locations in Two Streams in the Black Hills, South Dakota, USA

This study examined the spatial and temporal characteristics of brook trout Salvelinus fontinalis and brown trout Salmo trutta redds in two sites from two creeks in the Black Hills of South Dakota, USA. Spawning began in both streams in early October and continued through mid-November. Significant spatial clustering was only observed on both Rapid Creek sites, where-as random redd development was observed in both of the sites on Box Elder Creek. Based on visual observations, brown trout redds were more abundant in Rapid Creek, where-as brook trout redds were more abundant in Box Elder Creek. Differences in redd clustering could be due to species-specific, geological, or hydrological differences between the

Neurolin expression in the optic nerve and immunoreactivity of Pax6-positive niches in the brain of rainbow trout(Oncorhynchus mykiss) after unilateral eye injury

In contrast to astrocytes in mammals, fish astrocytes promote axon regeneration after brain injury and actively participate in the regeneration process. Neurolin, a regeneration-associated, Zn8-1abeled protein, is involved in the repair of damaged optic nerve in goldfish. At 1 week after unilateral eye injury, the ex- pression of neurolin in the optic nerve and chiasm, and the expression of Pax6 that influences nervous system development in various brain regions in the rainbow trout （Oncorhynchus mykiss） were detected. Immunohistochemical staining revealed that the number of Zn8~ cells in the optic nerve head and intraor- bital segment was obviously increased, and the increase in Zn8~ cells was also observed in the proximal and distal parts of injured optic nerve. This suggests that Zn8＊ astrocytes participate in optic nerve regeneration. ELISA results revealed that Pax6 protein increased obviously at 1 week post-injury. Immunohistochemi- cal staining revealed the appearance of Pax6＋ neurogenic niches and a larger number of neural precursor cells, which are mainly from Pax6＋ radial glia cells, in the nuclei of the diencephalon and optic tectum of rainbow trout （Oncorhynchus mykiss）. Taken together, unilateral eye injury can cause optic nerve reaction, and the formation of neurogenic niches is likely a compensation phenomenon during the repair process of optic nerve injury in rainbow trout （Oncorhynchus mykiss）.