Divergence in physiological factors affecting swimming performance between anadromous and resident populations of brook charr Salvelinus fontinalis.

In this study, an anadromous strain (L) and a freshwater-resident (R) strain of brook charr Salvelinus fontinalis as well as their reciprocal hybrids, were reared in a common environment and submitted to swimming tests combined with salinity challenges. The critical swimming speeds (Ucrit ) of the different crosses were measured in both fresh (FW) and salt water (SW) and the variations in several physiological traits (osmotic, energetic and metabolic capacities) that are predicted to influence swimming performance were documented. Anadromous and resident fish reached the same Ucrit in both FW and SW, with Ucrit being 14% lower in SW compared with FW. The strains, however, seemed to use different underlying strategies: the anadromous strain relied on its streamlined body shape and higher osmoregulatory capacity, while the resident strain had greater citrate synthase (FW) and lactate dehydrogenase (FW, SW) capacity and either greater initial stores or more efficient use of liver (FW, SW) and muscle (FW) glycogen during exercise. Compared with R♀ L♂ hybrids, L♀ R♂ hybrids had a 20% lower swimming speed, which was associated with a 24% smaller cardio-somatic index and higher physiological costs. Thus swimming performance depends on cross direction (i.e. which parental line was used as dam or sire). The study thus suggests that divergent physiological factors between anadromous and resident S. fontinalis may result in similar swimming capacities that are adapted to their respective lifestyles.

How does salinity influence habitat selection and growth in juvenile American eels Anguilla rostrata?

The influence of salinity on habitat selection and growth in juvenile American eels Anguilla rostrata captured in four rivers across eastern Canada was assessed in controlled experiments in 2011 and 2012. Glass eels were first categorized according to their salinity preferences towards fresh (FW), salt (SW) or brackish water (BW) and the growth rate of each group of elvers was subsequently monitored in controlled FW and BW environments for 7 months. Most glass eels (78-89%) did not make a choice, i.e. they remained in BW. Salinity preferences were not influenced by body condition, although a possible role of pigmentation could not be ruled out. Glass eels that did make a choice displayed a similar preference for FW (60-75%) regardless of their geographic origin but glass eels from the St Lawrence Estuary displayed a significantly higher locomotor activity than those from other regions. Neither the salinity preferences showed by glass eels in the first experiment nor the rearing salinities appeared to have much influence on growth during the experiments. Elvers from Nova Scotia, however, reached a significantly higher mass than those from the St Lawrence Estuary thus supporting the hypothesis of genetically (or epigenetically) based differences for growth between A. rostrata from different origins. These results provide important ecological knowledge for the sustained exploitation and conservation of this threatened species.

Prey exoskeletons influence the course of gastric evacuation in Atlantic cod Gadus morhua.

This study examined the effects of prey exoskeleton characteristics on gastric evacuation patterns in Atlantic cod Gadus morhua. Three distinct stages were highlighted in the gastric evacuation of crustacean prey characterized by a robust exoskeleton. The experiments confirmed that the three shrimp species, Pandalus borealis, Pandalus montagui and Eualus macilentus, and the crab Chionoecetes opilio, were evacuated from the stomach at different rates. The duration of all stages increased with increasing ash (and carbonate) content of the fresh prey. Thickness, chemical composition and morphology of the prey exoskeleton all affected gastric evacuation: duration of initial delay, overall evacuation rate and a decreased evacuation rate at the end of the process. The power exponential function (PEF), with its shape parameter, described the course of evacuation for these prey types well, especially the initial delay. The PEF does not, however, allow describing evacuation by the current stomach content mass independent of meal size, which limits its usefulness in estimating consumption rates of wild G. morhua. To predict and describe gastric evacuation of prey with a robust exoskeleton, it is therefore suggested that the square-root function be expanded with an initial lag phase, coupled to the mechanistically based cylinder model of gastric evacuation.

The influence of parental effects on transcriptomic landscape during early development in brook charr (Salvelinus fontinalis, Mitchill).

Parental effects represent an important source of variation in offspring phenotypes. Depending on the specific mechanisms involved, parental effects may be caused to different degrees by either the maternal or the paternal parent, and these effects may in turn act at different stages of development. To detect parental effects acting on gene transcription regulation and length phenotype during ontogeny, the transcriptomic profiles of two reciprocal hybrids from Laval × Rupert and Laval × Domestic populations of brook charr were compared at hatching, yolk sac resorption and 15 weeks after exogenous feeding. Using a salmonid cDNA microarray, our results show that parental effects modulated gene expression among reciprocal hybrids only at the yolk sac resorption stage. In addition, Laval × Domestic and Laval × Rupert reciprocal hybrids differed in the magnitude of theses parental effects, with 199 and 630 differentially expressed transcripts, respectively. This corresponds to a maximum of 18.5% of the analyzed transcripts. These transcripts are functionally related to cell cycle, nucleic acid metabolism and intracellular protein traffic, which is consistent with observed differences associated with embryonic development and growth differences in other fish species. Our results thus illustrate how parental effects on patterns of gene transcription seem dependent on the genetic architecture of the parents. In addition, in absence of transcriptional differences, non-transcript deposits in the yolk sac could contribute to the observed length differences among the reciprocal hybrids before yolk sac resorption.

Linking transcriptomic and genomic variation to growth in brook charr hybrids (Salvelinus fontinalis, Mitchill).

Hybridization can lead to phenotypic differences arising from changes in gene expression patterns or new allele combinations. Variation in gene expression is thought to be controlled by differences in transcription regulation of parental alleles, either through cis- or trans-regulatory elements. A previous study among brook charr hybrids from different populations (Rupert, Laval, and domestic) showing distinct length at age during early life stages also revealed different patterns in transcription regulation inheritance of transcript abundance. In the present study, transcript abundance using RNA-sequencing and quantitative real-time PCR, single-nucleotide polymorphism (SNP) genotypes and allelic imbalance were assessed in order to understand the molecular mechanisms underlying the observed transcriptomic and differences in length at age among domestic × Rupert hybrids and Laval × domestic hybrids. We found 198 differentially expressed genes between the two hybrid crosses, and allelic imbalance could be analyzed for 69 of them. Among these 69 genes, 36 genes exhibited cis-acting regulatory effects in both of the two crosses, thus confirming the prevalent role of cis-acting regulatory elements in the regulation of differentially expressed genes among intraspecific hybrids. In addition, we detected a significant association between SNP genotypes of three genes and length at age. Our study is thus one of the few that have highlighted some of the molecular mechanisms potentially involved in the differential phenotypic expression in intraspecific hybrids for nonmodel species.

Quantitative genetic analysis of the physiological stress response in three strains of brook charr Salvelinus fontinalis and their hybrids.

Three strains [domestic (D), Laval (L) and Rupert (R)] of brook charr Salvelinus fontinalis and their reciprocal hybrids were submitted to transport stress to measure stress resistance. Primary (cortisol) and secondary (glucose, osmolality and haematocrit) stress responses were measured for each cross. Significant heritabilities were observed for both levels of stress response, with mean ± S.E. heritability (h(2)) = 0.60 ± 0.20 for plasma cortisol and 0.61 ± 0.20 for plasma glucose. There were strain differences whereby the R strain was the least sensitive to stress at the primary and secondary levels. No heterosis was detected, and only one case of outbreeding depression was present. The outbreeding depression was observed in the D(♀) R(♂) hybrid, which had a 27% increase of plasma glucose compared to parental strains. The D(♀) R(♂) and R(♀) L(♂) hybrids had more pronounced variations (increase or decrease) in plasma osmolality than their respective parental strains, but these variations were difficult to relate definitively with the potential secondary stress response. These results indicate a strong potential for genetic improvement in the stress response to transport with the use of purebred crosses while hybridization has little value in this regard.

Poster - Thurs Eve-06: Maximizing eclipse IMRT dose accuracy by adjusting the dosimetric leaf gap parameter.

The dosimetric leaf gap (DLG) is a parameter used by Eclipse to model the rounded leaf ends of Varian MLCs. The DLGs were determined for the Millennium (M120) and High-Definition (HD120) model MLCs and taken as the difference between measured (0.6mm diode, IBA) and nominal MLC-defined profile FWHM values. Configuring the Eclipse pencil beam algorithm with the measured DLG gave poor agreement between measured and calculated IMRT dose distributions for the HD120 but not the M120. Agreement was optimized by adjusting the DLG for the HD120; 0.3mm changes in DLG were enough to cause significant variations in field dose agreement. Optimal DLG values of 0.04cm and 0.05cm were found for the 6MV HD120 and 10MV HD120, respectively, and 0.135cm 0.175cm for the 6MV M120 and 18MV M120, respectively. Agreement between measured and calculated dose distributions worsened for the AAA algorithm indicating separate DLG values may be required. A leaf calibration software upgrade also reduced agreement by changing the physical leaf position for a given location value. The change was detected using film and the picket fence MLC-pattern which places the two banks of opposing leaves at the same position but at different times. The DLG value can be adjusted from its measured physical value to improve the dosimetric accuracy of Eclipse IMRT plans and compensate for the effects of treatment planning algorithm and varying leaf calibrations. Since leaf calibrations are variable it is important to define the dosimetric leaf gap for each accelerator and clinic.

Maternal genetic effects on adaptive divergence between anadromous and resident brook charr during early life history.

The importance of directional selection relative to neutral evolution may be determined by comparing quantitative genetic variation in phenotype (Q(ST)) to variation at neutral molecular markers (F(ST)). Quantitative divergence between salmonid life history types is often considerable, but ontogenetic changes in the significance of major sources of genetic variance during post-hatch development suggest that selective differentiation varies by developmental stage. In this study, we tested the hypothesis that maternal genetic differentiation between anadromous and resident brook charr (Salvelinus fontinalis Mitchill) populations for early quantitative traits (embryonic size/growth, survival, egg number and developmental time) would be greater than neutral genetic differentiation, but that the maternal genetic basis for differentiation would be higher for pre-resorption traits than post-resorption traits. Quantitative genetic divergence between anadromous (seawater migratory) and resident Laval River (Québec) brook charr based on maternal genetic variance was high (Q(ST) > 0.4) for embryonic length, yolk sac volume, embryonic growth rate and time to first response to feeding relative to neutral genetic differentiation [F(ST) = 0.153 (0.071-0.214)], with anadromous females having positive genetic coefficients for all of the above characters. However, Q(ST) was essentially zero for all traits post-resorption of the yolk sac. Our results indicate that the observed divergence between resident and anadromous brook charr has been driven by directional selection, and may therefore be adaptive. Moreover, they provide among the first evidence that the relative importance of selective differentiation may be highly context-specific, and varies by genetic contributions to phenotype by parental sex at specific points in offspring ontogeny. This in turn suggests that interpretations of Q(ST)-F(ST) comparisons may be improved by considering the structure of quantitative genetic architecture by age category and the sex of the parent used in estimation.

CT gel dosimetry technique: comparison of a planned and measured 3D stereotactic dose volume.

This study presents a 3D dose mapping of complex dose distributions using an x-ray computed tomography (CT) polymer gel dosimetry technique. Two polyacrylamide gels (PAGs) of identical composition were irradiated with the same four arc stereotactic treatment to maximum doses of 15 Gy (PAG1) and 8 Gy (PAG2). The PAGs were CT imaged using a previously defined protocol that involves image averaging and background subtraction to improve image quality. For comparison with the planned isodose distribution, the PAG images were converted to relative dose maps using a CT number-dose calibration curve or simple division. The PAG images were then co-registered with the planning CT images in the BrainLab treatment planning software which automatically provides reconstructed sagittal and coronal images for 3D evaluation of measured and planned dose. The hypo-intense high dose region in both sets of gel images agreed with the planned 80% isodose contour and was shifted by up to 1.5 and 3.0 mm in the axial and reconstructed planes, respectively. This demonstrates the ability of the CT gel technique to accurately localize the high dose region produced by the stereotactic treatment. The resulting agreement of the measured relative dose volume for PAG1 was within 3.0 mm for the 50% and 80% isodose surfaces. However, the dose contrast was too low in PAG2 to allow for accurate definition of measured relative dose surfaces. Thus, a PAG should be irradiated to higher doses if quantitative relative dose information is required. Unfortunately, this implies use of an additional PAG and its CT number dose response since doses greater than 8-10 Gy fall outside the linear regions of the response.

Characterization of monomer/crosslinker consumption and polymer formation observed in FT-Raman spectra of irradiated polyacrylamide gels.

Fourier transform Raman spectroscopy was undertaken in the study of irradiated polyacrylamide gels (PAGs) used in 3D radiation dosimetry. By employing correlation techniques, monomer and crosslinker consumption were characterized in the spectra as a function of absorbed dose. The consumption of both monomer and crosslinker is monoexponential up to 13 Gy, although the rates of consumption differ for the two molecules. A sensitivity parameter, D0, in the exponential function has been used to characterize this difference. Up to 13 Gy, D0(acr) = 12 +/- 2 Gy while D0(bis) = 8.0 +/- 0.5 Gy, indicating that bis is consumed at a greater rate than acrylamide and that bis is the limiting factor in the onset of gel saturation, for a gel composition of 6% by weight total monomer (6%T) and where 3% of the total monomer is crosslinker (50%C). Direct evidence of polymer formation was observed in the Raman spectra of irradiated PAG. Polymer formation is monoexponential to a dose of 13 Gy, with a sensitivity parameter of D0(poly) = 14 +/- 2 Gy. This is in good agreement with the consumption rate of acrylamide. The exponential nature of the polymer formation observed here is compared with existing MRI and x-ray CT dose response measurements previously reported to be linear. The results confirm previous studies indicating that Raman spectroscopy provides a direct and useful tool for characterization of irradiated PAG.

Polymer gel dosimetry using x-ray computed tomography: a feasibility study.

A new three-dimensional dosimetry technique using x-ray computed tomography (CT) to analyse polymer gels is proposed. The CT imaging is sensitive to radiation-induced density changes that occur within irradiated polyacrylamide gel (PAG). In this preliminary study, a CT imaging protocol is developed to optimize CT images of PAG; the response of PAG CT number to dose (N(CT)-dose response) and the reproducibility of the response are investigated, and the use of CT to analyse PAG is compared with MRI. Experiments were conducted using two 1.5 l cylindrical PAG phantoms (3% acrylamide, 3% bis and 5% gelatin by weight), one irradiated with four intersecting 10 MV photon beams and the other with 10 sets of 6 MV parallel opposed circular radiosurgery fields. The final imaging protocol involves using optimum CT parameters (120 kVp and 200 mAs for our GE HiSpeed CT/i scanner), image averaging and background subtraction. The N(CT)-dose response is reproducible, linear up to 800-1000 cGy and is relatively insensitive to the gel temperature during imaging. The dose resolution is approximately 50 cGy for an image thickness of 10 mm. Despite the low dose resolution, preliminary results indicate that this CT technique provides accurate localization of high dose gradients such as those observed in stereotactic radiosurgery. Thus, given the availability and speed of CT scanners, the technique has the potential to be a valuable and practical 3D dose verification tool in radiation therapy.

Effects of long-term temperature acclimation on thyroid hormone deiodinase function, plasma thyroid hormone levels, growth, and reproductive status of male Atlantic cod, Gadus morhua.

The recent collapse of the Northwestern Atlantic cod fisheries has coincided with a cooling of water temperatures. During this time the condition factor of cod has been poor. The objective of the present study was to determine the effects of long-term temperature acclimation on growth reproduction and thyroid function in laboratory held Atlantic cod (Gadus morhua). One of the key parameters used to assess thyroid function is the peripheral metabolism of L-thyroxine (T4) by microsomal deiodinase enzymes. Deiodinase function has not been described for gadid fish. T4 outer-ring deiodinating activity (apparent K(m) 1-2 nM) was confined primarily to liver. Its properties resembled those for hepatic T4ORD activity of other teleosts and the mammalian type II deiodinase. The T4ORD activity of cod liver exceeded that of salmonids and could explain the high plasma T3 levels (10-18 ng/ml), which were 2-5 times greater than T4 levels. T4 and T3 inner-ring deiodination was confined mainly to brain. In order to determine the effects of long-term temperature acclimation on cod, somatic growth, reproduction, and thyroidal status were assessed monthly in 400-900-g satiation-fed male Atlantic cod captured in June from the St. Lawrence Estuary and then acclimated from August to the following June under a natural photoperiod at 2-4 degrees C (LT) or 6-10 degrees C (HT). Reproductive status was determined from the gonadosomatic index (GSI), plasma testosterone (T) and 11-ketotestosterone (11-KT) levels, and the appearance of milt; thyroidal status was determined from plasma T4 and 3,5,3'-triiodo-L-thyronine (T3) levels and hepatic T4ORD activity to produce biologically active T3. Testis maturation (high levels of 1 and 11-KT, and milt release) occurred in April and May and was uninfluenced by acclimation temperature. LT cod grew more slowly than HT cod. Differences in body weight were particularly evident from December to February. In conclusion, (i) cod possess outer- and inner-ring deiodinase activities, predominating respectively in liver and brain, and with properties resembling those of other teleosts, (ii) T4ORD activity of liver is unusually high and may account for the high plasma T3 levels in this species, (iii) T4ORD activity tends to increase during periods of increased somatic growth, and (iv) chronic acclimation of male cod to 2-4 degrees C, as opposed to 6-10 degrees C, decreases somatic growth but does alter circulating levels of thyroid hormones and androgens and it does not change the time of sexual maturation.

Effects of crosslinking and temperature on the dose response of a BANG polymer gel dosimeter.

The effects of varying the weight fraction (%C) of the crosslinker N, N'-methylene-bisacrylamide (bis) per total amount of monomer (6% w/w), and the NMR measurement temperature, on the dose response of the transverse relaxation rate (R2) of bis-acrylamide-nitrogen-gelatin (BANG) aqueous polymer gel dosimeters have been investigated. The gel samples were irradiated in test tubes with 250 kV x-rays, and the water proton NMR transverse relaxation rates were measured at 0.47 T using a Carr-Purcell-Meiboom-Gill multiecho pulse sequence. Both the dose sensitivity (slope of the linear portion of an R2-dose response) and the maximum rate at which the R2-dose response saturated (R2max), were found to depend strongly on the crosslinker fraction and on the temperature of the R2 measurement. The dose sensitivity peaked at approximately 50% C, and, for this composition, varied from 0.14 s-1 Gy-1 at 40 degrees C to 0.48 s-1 Gy-1 at 10 degrees C. The maximum transverse relaxation rates ranged from 0.8 s-1 at 33% C and 40 degrees C to 11.8 s-1 at 83% C and 5 degrees C. These results suggest that water proton transverse relaxation in the gel is controlled by an exchange of magnetization between the aqueous phase and the semi-solid protons associated with the polymer, and that the latter experience spectral broadening from immobilization which increases with crosslinking or cooling. Theoretical and practical implications of the above findings are discussed in the paper.

Multiple-site fast exchange model for spin-lattice relaxation in the Fricke-gelatin dosimeter.

The ferrous sulfate-doped gel dosimeters have been developed for three-dimensional magnetic resonance imaging of radiation dose distributions. When the gel dosimeter is irradiated, ferrous ions are converted to ferric ions and the nuclear magnetic spin relaxation of the dosimeter varies with dose. In this paper, a model is presented for the dose dependence of the spin-lattice relaxation rate R1 of the ferrous sulfate doped-gelatin dosimeter. The model is based on three basic physical quantities: the ferric ion yield and the ferrous and ferric ion relaxivities, r2+ and r3+, respectively. These relaxivities specify the ability of the ions to enhance the spin-lattice relaxation of water protons. The effects of gelatin and sulfuric acid concentration on the ferric ion yield and ion relaxivities are presented. The measured r2+ values agree with those predicted by a model in which the measured spin relaxation is considered the result of the fast exchange of water hydrating the ferrous ion with water in the bulk. The r3+ values are lower than predicted by the fast exchange model. The discrepancies in the measured and predicted r3+ values are shown to result from the complexing of ferric ions arising from pH variation caused by changes in gelatin or sulfuric acid concentrations. A modified version of the R1-dose response model accounting for ferric ion complexing is presented and tested spectrophotometrically.

Impaired cortisol stress response in fish from environments polluted by PAHs, PCBs, and mercury.

The cortisol stress response to capture was investigated in two species of fish (Perca flavescens and Esox lucius) from sites polluted by high levels of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and mercury, and from reference sites in the St. Lawrence river system. Fish from the reference sites exhibited the normal elevation of serum cortisol in response to the acute stress of capture and had large pituitary corticotropes. In contrast, fish from the most polluted sites were unable to increase their serum cortisol in response to the acute stress of capture and their pituitary corticotropes were atrophied. These results suggest that a life-long exposure to chemical pollutants may lead to an exhaustion of the cortisol-producing endocrine system, possibly as a result of prolonged hyperactivity of the system.

Influence of light on the ferredoxin-dependent glutamate synthase in maize leaves.

The ferredoxin (Fd)-dependent glutamate synthase (EC 1.4.7.1) and NADH-dependent glutamate synthase (EC 1.4.1.14) activities are carried out by two immunochemically distinct enzyme proteins in maize leaves (Zea mays W64A and W182E). Continuous irradiation of etiolated tissue at 75 micro einsteins per square meter per second for 24 hours resulted in a 3-fold increase on a fresh weight basis in the activity of the Fd-dependent glutamate synthase and a slight decrease in the activity of the NADH-dependent enzyme. There was also a significant increase of the Fd-glutamate synthase protein during greening of etiolated tissue.

Photoperiod effects on plasma cortisol levels in Gasterosteus aculeatus.

No diel variations of plasma cortisol levels were found in Gasterosteus aculeatus acclimated to two different photoperiod conditions: 14L:10D and 9L:15D. However, under long photoperiods, G. aculeatus exhibited higher plasma cortisol levels than under short photoperiods, suggesting that elevated plasma cortisol levels are associated with the spring migration and/or reproductive activities. This decrease of plasma cortisol levels is particularly marked in male G. aculeatus. Under the 9L:15D photoperiod, males had lower plasma cortisol levels than females, in contrast to 14L:10D, where no significant sex differences were found.