Kocuria SM1 controls vibriosis in rainbow trout (Oncorhynchus mykiss,Walbaum)

Aims: To develop probiotics for the control of vibriosis caused by Vibrio anguillarum and Vibrio ordalii in finfish. Methods and Results: Kocuria SM1, isolated from the digestive tract of rainbow trout, was administered orally to rainbow trout (Oncorhynchus mykiss) for 2 weeks at a dose equivalent to c. 10⁸ cells per g of feed and then challenged intraperitoneally with V. anguillarum and V. ordalii. Use of SM1 led to a reduction in mortalities to 15–20% compared to 74–80% mortalities in the controls. SM1 stimulated both cellular and humoral immune responses in rainbow trout, by elevation of leucocytes (5·5 ± 0·8 × 10⁶ ml^?1 from 3·7 ± 0·8 × 10⁶ ml^?1), erythrocytes (1·2 ± 0·1 × 10⁸ ml^?1 from 0·8 ± 0·1 × 10⁸ ml^?1), protein (23 ± 4·4 mg ml^?1 from 16 ± 1·3 mg ml^?1), globulin (15·7 ± 0·2 mg ml^?1 from 9·9 ± 0·1 mg ml^?1) and albumin (7·3 ± 0·2 mg ml^?1 from 6·1 ± 0·1 mg ml^?1) levels, upregulation of respiratory burst (0·05 ± 0·01 from 0·02 ± 0·01), complement (56 ± 7·2 units ml^?1 from 40 ± 8·0 units ml^?1), lysozyme (920 ± 128·8 units ml^?1 from 760 ± 115·3 units ml^?1) and bacterial killing activities. Conclusions: Kocuria SM1 successfully controlled vibriosis in rainbow trout, and the mode of action reflected stimulation of the host innate immune system. Significance and Impact of the Study: Probiotics can contribute a significant role in fish disease control strategies, and their use may replace some of the inhibitory chemicals currently used in fish farms.     

Microbial diversity of intestinal contents and mucus in rainbow trout (Oncorhynchus mykiss)

AIMS: The aim of this study was to understand the microbial community of intestinal contents and mucosal layer in the intestine of rainbow trout by means of culture-dependent conventional and independent molecular techniques. METHODS AND RESULTS: Forty-one culturable microbial phylotypes, and 39 sequences from 16S rRNA and two from 18S rRNA genes, were retrieved. Aeromonadaceae, Enterobacteriaceae and Pseudomonadaceae representatives were the dominant cultured bacteria. Genomic DNA isolated from intestinal contents and mucus was used to generate 104 random clones, which were grouped into 32 phylotypes at 99% minimum similarity, most of which were affiliated with Proteobacteria (>70% of the total). However, unlike library C (intestinal contents), the phyla Bacteroidetes and Fusobacteria were not found in intestinal mucus (library M), indicating that the microbiota in the gut mucus was different from that of the intestinal contents. Twelve sequences were retrieved from denaturing gradient gel electrophoresis analysis, and dominant bands were mostly related to Clostridium. CONCLUSIONS: Many novel sequences that have not been previously recognized as part of the intestinal flora of rainbow trout were retrieved. SIGNIFICANCE AND IMPACT OF THE STUDY: The fish gut harbours a larger bacterial diversity than previously recognized, and the diversity of gut mucus is different from that of intestinal contents.     

溶解氧对虹鳟呼吸机能及红细胞特性的影响

目的：初步研究溶解氧对虹鳟呼吸机能及红细胞特性的影响。方法：设四组溶氧水平,测定呼吸频率、血液中PO2、PCO2和pH值,并以RBC、Hb、脆性、膜粘滞性及丙二醛含量等指标观察溶氧对红细胞的影响。结果：溶解氧升高使鱼呼吸频率降低,血液PO2升高（P〈0.05）;高溶氧导致红细胞数、丙二醛含量降低。红细胞膜粘滞系数在试验第10d时,高氧组明显高于其它组,但30d时各溶氧组差异不显著。结论：在一定范围内溶解氧升高可以改善机体组织氧含量,提高呼吸功能,红细胞对溶氧变化有其生理适应性。     

Enhancement of the immune response and protection induced by probiotic lactic acid bacteria against furunculosis in rainbow trout (Oncorhynchus mykiss)

We analysed the effect of probiotic strains on the cellular and humoral immune responses of rainbow trout (Oncorhynchus mykiss), and their capacity to prevent furunculosis during a challenge trial. Probiotic strains (Lactococcus lactis ssp. lactis CLFP 100, Leuconostoc mesenteroides CLFP 196, and Lactobacillus sakei CLFP 202) were administered orally to fish for 2 weeks at 10(6) CFU g(-1) of feed. In comparison to untreated control fish, the phagocytic activity of head kidney leukocytes and the alternative complement activity in serum were significantly greater in all probiotic groups at the end of the second week. With the exception of the group fed with Lactobacillus sakei, superoxide anion production was also significantly increased in the probiotic groups. Analysis of lysozyme activity did not exhibit any significant difference in the probiotic and control groups. Fifteen days after the start of the probiotic feeding, fish were challenged with Aeromonas salmonicida ssp. salmonicida. The fish supplemented with probiotics exhibited survival rates ranging from 97.8% to 100%, whereas survival was 65.6% in fish not treated with the probiotics. These results demonstrate that probiotic supplementation to fish can reduce the severity of furunculosis, and suggest that this reduction may be associated with enhanced humoral and cellular immune response.     

Effects of nonylphenol on hematological parameters and immune responses in immature rainbow trout (Oncorhynchus mykiss)

ABSTRACT

This study investigated the effects of nonylphenol (NP) on hematological and immunological parameters in both male and female rainbow trout (Oncorhynchus mykiss). Fish were randomly distributed into six groups and administered with NP (10, 50 and 100 μg g^-1 week^-1 BW) and a single dose of 17-β estradiol (E2; 2 μg g^-1 week^-1 BW, positive control). The solvent controls received ethanol and coconut oil as a vehicle, while the controls were not injected. Red blood cells (RBCs) count, hematocrit (Hct), hemoglobin (Hb), white blood cells (WBCs), and lymphocytes demonstrated a NP dose-dependent decrease, whereas mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), monocytes, and neutrophils showed an increasing trend in both male and female fish 21 days post-treatment. Also, RBCs, Hb, MCHC, WBCs, and lymphocytes were significantly reduced (p<0.05) in E2 treated fish. Lysozyme, complement components (C3 and C4) and immunoglobulin M (IgM) were increased in fish sera subjected to 10 and 50 μg g^-1 NP, while these decreased in groups administered with 100 μg g^-1 NP and 2 μg g^-1 E2. Except for C4 level at 10 μg g^-1 NP, no significant differences were observed in hematological and immunological parameters of male and female in each treatment. Overall, a frequent exposure to NP could lead to adverse effects on fish immune-physiological functions which may cause serious ecological threats of fish natural population sustainability.     

Cortisol and immune characteristics in rainbow trout (Oncorhynchus mykiss) selected for high or low tolerance to stress

The influence of exposure to stressors on cortisol and the non-specific immune traits lysozyme and serum haemolytic activity were examined in second generation rainbow trout ( Oncorhynchus mykiss ) selected for either high or low serum cortisol level following a confinement stress. Lysozyme and serum haemolytic activity were also assessed, together with levels of specific antibodies against Aeromonas salmonicida A-layer, Vibrio salmonicida O-antigen and Vibrio anguillarum O-antigen, following injection of vaccines against these pathogens.
Significant differences in mean cortisol levels between the two selection lines were observed, but in only one of two stress experiments was the 'high-stress' line found to have the higher cortisol level; in the other experiment the 'high-stress' line had significantly lower cortisol levels than the 'low-stress' line. Lysozyme levels were in four of four assessments higher in the high-stress line than in the low-stress line, whereas components of serum haemolytic activity tended to be lower in the high-stress line than in the low-stress line. Levels of specific antibodies against all three bacterial pathogens were elevated following the injection of the vaccines. Only antibody production against A. salmonicida A-layer was significantly different between the two lines, the higher production of antibody being in the high-stress line.     

Characterisation of mucosal and systemic immune responses in rainbow trout (Oncorhynchus mykiss) using surface plasmon resonance

Mucosal and systemic antibody production in rainbow trout, Oncorhynchus mykiss (Walbaum), was evaluated following different antigen delivery routes. A BIAcore instrument (Pharmacia) allowed direct detection of antibody-antigen interactions by surface plasmon resonance changes. These interactions were measured in real-time without secondary reagents or extraneous labels. Groups of rainbow trout were immunised with a hapten-carrier antigen consisting of fluorescein isothiocyanate (FITC) conjugated to keyhole limpet haemocyanin (KLH) or phosphate buffered saline (PBS) pH 7.2. Antigens were administered intraperitoneally (i.p.) with or without Freund's complete adjuvant (FCA) or peranally (p.a.) directly to the gastrointestinal (GI) tract. Serum and mucosal anti-FITC responses were significantly (P<0.05) higher in FITC-KLH/FCA groups, clearly showing that adjuvant incorporation enhances mucosal as well as sytemic immunity. Antigen uptake and processing in fish immunised p.a. and i.p. without adjuvant was much less efficient and resulted in relatively low levels of serum and mucosal antibody production. Interestingly, mucosal responses in these groups peaked prior to serum responses suggesting possible early stimulation of mucosal defences. Mucosal antibody production in fish receiving FITC-KLH/FCA correlated more closely with serum responses, indicating possible transfer of serum derived antibody to mucosal sites. Mucosal and serum responses were confirmed as immunoglobulin (Ig) by subsequent reactivity with an anti-trout serum IgM monoclonal antibody (1.14) passed over flow cells containing anti-FITC antibodies. Further analysis showed significantly lower (P<0.05) reactivity of early mucus anti-FITC components (4 weeks post-immunisation) to 1.14. Purified serum and mucus Ig from non-immunised fish showed different protein banding patterns by SDS-PAGE under reducing conditions. Immunoblotting with 1.14 also showed weak reactivity to mucus Ig in control fish while reacting strongly to mucus Ig from immunised fish. These data suggest that early mucosal responses in trout may consist of heterogeneous forms of Ig differing in characteristics to serum Ig. BIAcore analysis in this context and as a means of measuring antibody response proved useful, and has the potential to become a valuable new tool in the study of fish immunology.     

Effects of genetic sex and genomic background on epistasis in rainbow trout (Oncorhynchus mykiss)

Epistasis among quantitative trait loci (QTL) for survival (upper thermal tolerance, UTT) and morphological (fork length, FL and condition factor, K) traits was detected in purestrain and interstrain rainbow trout (Oncorhynchus mykiss) families. One sex-linked (OmyFGT19TUF) and three autosomal (Omy325UoG, Ssa14DU and Ssa20.19NUIG; linkage groups B, D and S, respectively) microsatellite loci linked to UTT QTL in this species were used. Within half sib families, significant effects of full sib family on epistasis involving Omy325UoG and OmyFGT19TUF were detected at a rate significantly higher than expected for UTT (p < 0.001*) and FL (p < 0.01*), using results significant at comparisonwise significance thresholds derived from permutational analysis. Measured across half sib families, the phenotype of female genotypic classes was more divergent from the family trait mean than that of males where epistasis involved the sex-linked locus OmyFGT19TUF (p = 0.0176*), and also for means over all families (p = 0.0355*). Female genotypic classes were also more divergent (p = 0.0011**) from the full sib trait mean where three-way interaction between OmyFGT19TUF, one of the autosomal loci and full sib family was significant, and marginally more divergent for trait means of genotypic classes across all full sib families (p = 0.0856). There was no evidence that these effects were more pronounced in hybrid F₁ families than purestrains.     

Red beet and betaine as ingredients in diets of rainbow trout (Oncorhynchus mykiss): effects on growth performance,nutrient retention and flesh quality

The objective of the study was to evaluate the impact of different concentrations of dietary red beet and betaine on the growth performance and fish flesh quality of rainbow trout. Therefore, a control diet was compared with four diets in which two levels of red beet (14% and 28%) and betaine (0.9% and 1.63%) were incorporated in combination. The study was set up with an average body weight of 69 ± 2.2 g and finished when fish reached commercial weight (175–250 g) after 105 d. The impact of the diets was studied based on the growth performance, biometric indexes, proximal composition, protein and fat retention efficiencies and apparent nutrient digestibility by fish reared on a recirculation system. Further estimates were the effect of red beet and betaine on the flesh proximate composition and quality of the final product (water activity, colour, texture, thiobarbituric acid reactive substances and sensory characteristics). Results showed that inclusion of 14% red beet and 0.9% betaine did not affect growth, nutritive or biometric parameters and nutrient retention when compared with the control diet. However, higher levels of red beet and betaine had negative effects on growth and nutritive parameters. The tested ingredients enhanced quality parameters regardless of the concentration used. After feeding the red beet and betaine, fish flesh showed lower water activity and better textural and colour properties than the control and also a dose-dependent effect on lipid oxidation was observed.     

Analysis of the association between spawning time QTL markers and the biannual spawning behavior in rainbow trout (Oncorhynchus mykiss)

The rainbow trout is a salmonid fish that occasionally exhibits broodstocks with biannual spawning behavior, a phenomenon known as a double annual reproductive cycle (DARC). Spawning time quantitative trait loci (SPT-QTLs) affect the time of the year that female rainbow trout spawn and may influence expression of the DARC trait. In this study, microsatellite markers linked and unlinked to SPT-QTLs were genotyped to investigate the underlying genetics of this trait. SPT-QTLs influenced the DARC trait since in two case-control comparisons three linked markers (OmyFGT12TUF, One3ASC and One19ASC) had significant levels of allelic frequency differentiation and marker-character association. Furthermore, alleles of One3ASC and One19ASC had significantly higher frequencies in populations that carried the DARC trait.     

Primary and secondary indices of stress in the progeny of rainbow trout (Oncorhynchus mykiss) selected for high and low responsiveness to stress

A series of pooled gamete matings was carried out employing eggs and milt from mature male and female rainbow trout selected for a consistently high- or low-responsiveness to stress, as indicated by post-stress plasma cortisol elevation. Development of the progeny was closely monitored and the responsiveness to stress of the progeny of high-responding parents and the progeny of low-responding parents was assessed by two methods. For a period of 14 months, at approximately monthly intervals, the plasma cortisol elevation evoked by a standardized confinement stress was determined in fish from each group, and secondly, on one occasion, the time-course of the plasma cortisol response to a 24-h period of confinement was monitored. Progeny of high-responding parents snowed a significantly greater cortisol response to stress than the progeny of low-responding parents during both testing procedures. However, when the effect of a 14-day confinement stress was examined, high-responding progeny showed a more rapid recovery of plasma cortisol levels, while levels in the low-responding progeny, although initially lower, showed a more sustained elevation. To assess the possible functional implications of these observations, circulating lymphocyte numbers, an immunologically important cortisol-sensitive component of the blood cell complement, were determined. The duration of the lymphocytopenia observed following the onset of confinement was found to be related to the initial, not the sustained, cortisol response. These data suggest that manipulation of the sensitivity to stress of fish is feasible by selective breeding, but that careful.choice of the indices employed to identify traits considered desirable is necessary.     

Efficacy of in-feed probiotics against Aeromonas bestiarum and Ichthyophthirius multifiliis skin infections in rainbow trout (Oncorhynchus mykiss, Walbaum)

Aims:  The aim of this study was to assess the efficacy of in-feed probiotics as a preventive measure against skin infections caused by Aeromonas bestiarum and Ichthyophthirius multifiliis (Ich) in rainbow trout.
Methods and Results:  Fin rot was induced in fish by intradermal injection with 0·1 ml volumes containing 10⁵ cells per ml A. bestiarum at the base of the dorsal fin. Ich infections resulted from immersion in Ich-contaminated water. Each probiotic was administered orally [10⁸ cells per g feed for GC2 ( Aeromonas sobria ) and 10¹⁰ cells per g feed for BA211 ( Brochothrix thermosphacta )] for 14 days. Results showed that, after challenge with A. bestiarum , probiotics GC2 and BA211 led to 76% and 88% survival, respectively, in contrast to 22% survival for controls. Fish fed with probiotic GC2 had 100% survival after challenge with Ich compared with 2% for probiotic BA211 and 0% for controls. Analysis of innate immune responses revealed that probiotic GC2 promoted higher phagocytic activity, whereas probiotic BA211 led to enhanced respiratory burst activity.
Conclusion:  Of the two probiotics examined, GC2 was more effective in protecting against both fin rot and Ich. Each probiotic appeared to stimulate different pathways within the innate immune system.
Significance and Impact of the Study:  This is the first demonstration that probiotics can protect fish against surface infections. Furthermore, this is the first time a probiotic has been shown to protect against a eucaryotic pathogen, namely I. multifiliis .     

Liver and muscle fatty acid composition of mature and immature rainbow trout (<Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>) fed two different diets

Fatty acid composition of liver and muscle tissues of immature and mature Oncorhynchus mykiss fed on two different diets were determined. Fatty acid analyses were carried out by gas chromatography. Palmitic acid (C16:0), oleic acid (C18:1 n-9), linoleic acid (C18:2 n-6) and docosahexaenoic acid (C22:6 n-3) were the major components in both liver and muscle tissues of immature and mature rainbow trout of both sexes. The amounts of C22:6 n-3 were higher in the liver (29.04 ± 0.06 − 27.41 ± 0.17%) and muscle (13.05 ± 0.40 − 11.37 ± 0.21%) of immature fish than in mature fish and depended on the composition of the diet. Results of this study show that fatty acid composition in fish tissues can considerably vary, depending on the age of fish and their diet. Thus more detailed studies are necessary on the influence of diet on immature and mature fish fatty acid composition. The age and diet of fish consumed may also be important for human health.     

Effects of dehulling,steam-cooking and microwave-irradiation on digestive value of white lupin (Lupinus albus) seed meal for rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar)

A digestibility trial was conducted to assess the effect of dehulling, steam-cooking and microwave-irradiation on the apparent digestibility of nutrients in white lupin (Lupinus albus) seed meal when fed to rainbow trout (Oncorhynchus mykiss) and Atlantic salmon (Salmo salar). Six ingredients, whole lupin seed meal (LSM), dehulled LSM, dehulled LSM steam-cooked for 15 or 45 min (SC15 and SC45, respectively) and LSM microwave-irradiated at 375 or 750 W (MW375 and MW750, respectively), were evaluated for digestibility of dry matter, crude protein (CP), lipids, nitrogen-free extractives (NFE) and gross energy (GE). The diet-substitution approach was used (70% reference diet + 30% test ingredient). Faeces from each tank were collected using a settlement column. Dehulled LSM showed higher levels of proximate components (except for NFE and crude fibre), GE and phosphorus in comparison to whole LSM. Furthermore, SC15, SC45, MW375 and MW750 showed slight variations of chemical composition in comparison to dehulled LSM. Results from the digestibility trial indicated that dehulled LSM, SC15, SC45 and MW375 are suitable processing methods for the improvement of nutrients’ apparent digestibility coefficient (ADC) in whole LSM. MW750 showed a lower ADC of nutrients (except for CP and lipids for rainbow trout) in comparison with MW350 for rainbow trout and Atlantic salmon, suggesting a heat damage of the ingredient when microwave-irradiation exceeded 350 W.     

Plasma levels of gonadotropin and 17α,20β-dihydroxy-4-pregnen-3-one in relation to spawning behaviour of rainbow trout, Oncorhynchus mykiss(Walbaum)

Manipulation of the opportunity to spawn was used to investigate the relationship between endocrine events, egg viability and spawning behaviour in female rainbow trout. Females were prevented from spawning by isolating them from males and gravel for up to 21 days after ovula- tion. Blood samples were taken before pairing with a male, at the onset of nesting activity, and at the completion of spawning. Plasma hormone levels of gonadotropin (GtH) and 17α,20β-dihydroxy-4-pregnen-3-one (17,2OP) were measured by specific radioimmunoassays. There were no qualitative or quantitative differences in the spawning behaviour of females paired on the day of ovulation or 7. 14, or 21 days after ovulation. There was a general decrease in the viability of eggs with increasing retention times. In females paired on the day of ovulation, or after 7 or 14 days, GtH levels increased with the onset of nesting behaviour and declined as fish reached the post-spawning condition. By day 21, GtH levels before pairing were significantly higher than prepairing levels in the other three treatment groups, and did not increase at the onset of nesting, or decrease in post-spawning fish. Plasma 17,20P remained high in prepairing and nesting samples of all four groups and declined to low levels in fish in post-spawning condition. In females paired on the day of ovulation there was a significant increase in 17,20P from the prepairing to the nesting stage. These results suggest that 17,20P plays a key role in the synchronization of behavioural and maturational events at the time of spawning.     

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.     

Effect of fasting on thyroid hormone levels, and TR(alpha) and TR(beta) mRNA accumulation in late-stage embryo and juvenile rainbow trout, Oncorhynchus mykiss

The accumulation of mRNA encoding for hepatic and intestinal T3-receptor (TR) and body and liver masses were measured in fed and 3-week fasted juvenile and swim up stage rainbow trout embryos. Plasma and total body thyroid hormone (TH) levels were measured for juvenile and swim up stages, respectively. Fasted juveniles exhibited a lower hepatosomatic index (HSI), liver mass and plasma T4 and T3 concentrations than fed animals, but there were no changes in body mass or the accumulation of mRNA encoding for either of the TR(alpha) or TR(beta) isoforms in liver or intestine. TR(beta) mRNA accumulation was greater than TR(alpha) mRNA accumulation in both tissues. Fasted embryos had lower whole body TH levels and body, liver and intestinal tract masses, in addition to a lower intestinosomatic index. However, there was no change in HSI. Fasting did not affect whole body or hepatic TR(alpha) and TR(beta) mRNA accumulation, although intestinal tract TR(alpha) and TR(beta) mRNA accumulation was lower in the fasted embryos. The HSI and body mass changes in fasted juvenile and embryo stages, respectively, indicated that both developmental stages were impacted by fasting. Both stages also showed evidence of decreased TH production. The lower TR gene expression in the intestinal tract of fasted embryos may suggest a role for THs in the transitional stage of intestinal development during this period of development.     

Biomarker responses in juvenile rainbow trout (Oncorhynchus mykiss) after single and combined exposure to low doses of cadmium, zinc, PCB77 and 17β-oestradiol

The objective of this study was to examine (i) the biochemical responses of rainbow trout exposed to sublethal water concentrations of the metals cadmium (Cd) (1.5 μg l^-1) and zinc (Zn) (150 μg l^-1); and (ii) the potential combined effects when applied in mixture (Cd/Zn) with and without co-exposure to model organic chemicals 3,3',4,4'-tetrachlorobiphenyl (PCB77) (1 mg kg^-1) and 17β-oestradiol (E2) (0.5 mg kg^-1). After 21 days of exposure, several biomarkers were assessed in the liver (enzymatic and non-enzymatic antioxidants, heat shock proteins [HSP70 and HSP60], ethoxyresorufin-O-deethylase [EROD]) and in the plasma (vitellogenin [Vtg], aminotransferases). Plasma aminotransferases were not affected, whereas the other biomarkers showed different patterns of response depending on the treatment. For example, Cd, and Zn to a lesser extent, induced an adaptive response in the liver shown by an increase in antioxidant defences (total glutathione [GSH], superoxide dismutase, Trolox equivalent antioxidant capacity [TEAC]), without any impairment of GSH redox status or induction of heat shock proteins. Antagonistic effects were observed in GSH-related biomarkers after Cd/Zn exposure. PCB77 strongly induced EROD activity, HSP70 and TEAC. Co-exposure with metals did not modulate significantly the effects of PCB77. E2 induced Vtg and inhibited liver antioxidants and basal EROD activity. These inhibitory effects were suppressed in fishes exposed to E2 + Cd/Zn, suggesting additive effects of E2 and metals. In addition, E2-induced Vtg was not altered by metals. Multivariate analyses confirmed some correlation between the biomarkers. The use of complementary biomarkers is necessary to discriminate different treatments and to highlight interactive effects.     

Effects of cyclopenta[c]phenanthrene and its derivatives on zona radiata protein, ERalpha, and CYP1A mRNA expression in liver of rainbow trout (Oncorhynchus mykiss Walbaum)

Despite cyclopenta-fused polycyclic aromatic hydrocarbons (CP-PAHs) have been detected in the environment, the ability of CP-PAH to induce cellular and tissue responses remains poorly characterized. In this study, xenoestrogen-associated responses (mRNA levels of estrogen receptor alpha, ERalpha, and zona radiata protein, Zrp) and xenobiotic effects (CYP1A mRNA) have been investigated in liver of juvenile rainbow trout after short-term treatment (8 and 24 h) with following compounds administered singly: cyclopenta[c]phenanthrene (CP[c]Ph); its derivatives, 5A-CP[c]Ph; 5A6M-CP[c]Ph; 5A9M-CP[c]Ph; B[c]Ph, a structurally similar polycyclic aromatic hydrocarbon; B[a]P, a model CYP1A inducer; and zearalenone (ZEA), naturally occurring ligand for ER. The CYP1A mRNA expression after 24 h of exposure with CP[c]Ph or its derivatives, except 5A9M-CP[c]Ph, was 3-9-fold higher compared to controls (P<0.05), but it was less than that caused by B[a]P (65-fold up regulation; P<0.01). Moreover, neither of the CP-PAH compounds modulated liver ERalpha or Zrp mRNA levels as compared to effects associated with ZEA. Interestingly, a treatment with this ER-ligand, caused moderate but significant increase of CYP1A mRNA expression (about 2.5-fold; P<0.05). The finding that ZEA is capable of acting as either estrogenic and xenobiotic compound, should be further explored in a more detailed and differently designed experiment.     

Quantitative genetic and translocation experiments reveal genotype‐by‐environment effects on juvenile life‐history traits in two populations of Chinook salmon (Oncorhynchus tshawytscha)

Understanding the genetic architecture of phenotypic plasticity is required to assess how populations might respond to heterogeneous or changing environments. Although several studies have examined population‐level patterns in environmental heterogeneity and plasticity, few studies have examined individual‐level variation in plasticity. Here, we use the North Carolina II breeding design and translocation experiments between two populations of Chinook salmon to detail the genetic architecture and plasticity of offspring survival and growth. We followed the survival of 50 800 offspring through the larval stage and used parentage analysis to examine survival and growth through freshwater rearing. In one population, we found that additive genetic, nonadditive genetic and maternal effects explained 25%, 34% and 55% of the variance in larvae survival, respectively. In the second population, these effects explained 0%, 24% and 61% of the variance in larvae survival. In contrast, fry survival was regulated primarily by additive genetic effects, which indicates a shift from maternal to genetic effects as development proceeds. Fry growth also showed strong additive genetic effects. Translocations between populations revealed that offspring survival and growth varied between environments, the degree of which differed among families. These results indicate genetic differences among individuals in their degree of plasticity and consequently their ability to respond to environmental variation.