Céline Audet

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.

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 (QST) to variation at neutral molecular markers (FST). 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 (QST > 0.4) for embryonic length, yolk sac volume, embryonic growth rate and time to first response to feeding relative to neutral genetic differentiation [FST = 0.153 (0.071–0.214)], with anadromous females having positive genetic coefficients for all of the above characters. However, QST 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 QST‐FST comparisons may be improved by considering the structure of quantitative genetic architecture by age category and the sex of the parent used in estimation.

Network Analysis Highlights Complex Interactions between Pathogen, Host and Commensal Microbiota

Interactions between bacteria and their host represent a full continuum from pathogenicity to mutualism. From an evolutionary perspective, host-bacteria relationships are no longer considered a two-component system but rather a complex network. In this study, we focused on the relationship between brook charr (Salvelinus fontinalis) and bacterial communities developing on skin mucus. We hypothesized that stressful conditions such as those occurring in aquaculture production induce shifts in the bacterial community of healthy fish, thus allowing pathogens to cause infections. The results showed that fish skin mucus microbiota taxonomical structure is highly specific, its diversity being partly influenced by the surrounding water bacterial community. Two types of taxonomic co-variation patterns emerged across 121 contrasted communities’ samples: one encompassing four genera well known for their probiotic properties, the other harboring five genera mostly associated with pathogen species. The homeostasis of fish bacterial community was extensively disturbed by induction of physiological stress in that both: 1) the abundance of probiotic-like bacteria decreased after stress exposure; and 2) pathogenic bacteria increased following stress exposure. This study provides further insights regarding the role of mutualistic bacteria as a primary host protection barrier.

Shifting patterns in genetic control at the embryo-alevin boundary in brook charr

Abstract Maternal inputs to offspring early in development are initially high but the process of development suggests that ontogenetic shifts in the importance of maternal genetic variation relative to other sources should occur. We investigated additive genetic variance and covariance for direct (animal), sire, and maternal effects on embryonic length (EL), yolk sac volume (YSV), and alevin (after yolk sac resorption) length (AL) for 460 embryonic and 460 alevin brook charr (Salvelinus fontinalis) in 23 half‐sib families (12 sires, 23 dams). There were no additive genetic effects of sires or individual animals on their own phenotype using sire‐dam and maternal‐animal models for YSV or EL (h2 < 0.05). However, at the alevin stage we detected low but significant heritability for AL (h2= 0.14 ± 0.11). Conversely, maternal genetic effects were high for both embryonic traits (h2EL= 0.61 ± 0.05; h2YSV= 0.57 ± 0.06) but faded rapidly for postresorption length (h2AL= 0.18 ± 0.04). Maternal effects in the sire‐dam model corresponded highly with those in the animal‐dam model. We did not detect significant genetic covariance between progeny and dams for preresorption traits or between sires and dams for any trait. However, following resorption of the yolk sac, the genetic value of dams for AL was negatively correlated with that of individual progeny (rma=– 0.38 ± 0.13), suggesting trade‐offs and/or stabilizing selection between maternal and animal genetic trait value. This finding was supported by models of dam fecundity on offspring length and dam weight in phenotypic space. Heritability estimates using simple regression of embryo phenotype on adult parental phenotype produced upwardly biased estimates of genetic variance (h2± 1.0). We propose that development through the embryo‐alevin boundary may be a major point in salmonids for ontogenetic changes in the genetic architecture of embryo size from maternal genetic effects to those of the individual organism, nd that maternal‐offspring conflicts in resource allocation related to size may be partially indicated by negative genetic covariance.

The Transcriptional Landscape of Cross-specific Hybrids and its Possible Link with Growth in Brook Charr (Salvelinus fontinalis Mitchill)

The genetic mechanisms underlying hybridization are poorly understood despite their potentially important roles in speciation processes, adaptative evolution, and agronomical innovation. In this study, transcription profiles were compared among three populations of brook charr and their hybrids using microarrays to assess the influence of hybrid origin on modes of transcription regulation inheritance and on the mechanisms underlying growth. We found that twice as many transcripts were differently expressed between the domestic population and the two wild populations (Rupert and Laval) than between wild ones, despite their deeper genetic distance. This could reflect the consequence of artificial selection during domestication. We detected that hybrids exhibited strikingly different patterns of mode of transcription regulation, being mostly additive (94%) for domestic × Rupert, and nonadditive for Laval × domestic (45.7%) and Rupert × Laval hybrids (37.5%). Both heterosis and outbreeding depression for growth were observed among the crosses. Our results indicated that prevalence of dominance in transcription regulation seems related to growth heterosis, while prevalence of transgressive transcription regulation may be more related to outbreeding depression. Our study clearly shows, for the first time in vertebrates, that the consequences of hybridization on both the transcriptome level and the phenotype are highly dependent on the specific genetic architectures of crossed populations and therefore hardly predictable.

Inter individual variations of the fish skin microbiota: host genetics basis of mutualism?

The commensal microbiota of fish skin is suspected to provide a protection against opportunist infections. The skin of fish harbors a complex and diverse microbiota that closely interacts with the surrounding water microbial communities. Up to now there is no clear evidence as to whether the host regulates the recruitment of environmental bacteria to build a specific skin microbiota. To address this question, we detected Quantitative Trait Loci (QTL) associated with the abundance of specific skin microbiota bacterial strains in brook charr (Salvelinus fontinalis), combining 16S RNA tagged-amplicon 454 pyrosequencing with genetic linkage analysis. Skin microbiota analysis revealed high inter-individual variation among 86 F2 fish progeny based upon the relative abundance of bacterial operational taxonomic units (OTUs). Out of those OTUs, the pathogenic strain Flavobacterium psychrophilum and the non-pathogenic strain Methylobacterium rhodesianum explained the majority of inter-individual distances. Furthermore, a strong negative correlation was found between Flavobacterium and Methylobacterium, suggesting a mutually competitive relationship. Finally, after considering a total of 266 markers, genetic linkage analysis highlighted three major QTL associated with the abundance of Lysobacter, Rheinheimera and Methylobacterium. All these three genera are known for their beneficial antibacterial activity. Overall, our results provide evidence that host genotype may regulate the abundance of specific genera among their surface microbiota. They also indicate that Lysobacter, Rheinheimera and Methylobacterium are potentially important genera in providing protection against pathogens.

Physiological, endocrine, and genetic bases of anadromy in the brook charr, Salvelinus fontinalis, of the Laval River (Québec, Canada)

Brook charr, Salvelinus fontinalis, often display alternate life history styles in coastal areas. In the Laval River, some brook charr remain freshwater residents, while others undergo seasonal migrations between freshwater and saltwater environments. In the present paper, we examined physiological (electrolyte concentrations, gill Na+, K+-ATPase activity, and thyroid hormone levels) as well as genetic differences (neutral genetic markers) between anadromous and river-resident fish from the Laval River. We also examined how artificial rearing conditions affected seasonal variations in the osmoregulatory physiology of a domestic strain derived from wild anadromous fish. Sympatric anadromous and resident forms of brook charr of the Laval River exhibited differences in gill Na+, K+-ATPase activity, plasma thyroxine (T4), and triidothyronine (T3) concentrations. In domestic anadromous charr, rearing conditions during development had no negative impact on osmoregulatory ability or on gill Na+, K+-ATPase activity. These results argued for an important hereditary component of gill Na+, K+-ATPase activity. However, the spring increase in T4 was present only in wild fish. Significant differences observed at microsatellite loci further suggested that at least some level of reproductive isolation may have occurred between anadromous and resident charr in the Laval River.

Antagonistic effect of indigenous skin bacteria of brook charr (Salvelinus fontinalis) against Flavobacterium columnare and F. psychrophilum

Industrial fish production exposes fish to potentially stressful conditions, which in turn may induce infections by opportunistic pathogens. Probiotics appear to be a promising way to prevent opportunistic infections in aquaculture. In this study, we tested the inhibitory potential of endogenous bacterial communities found in the mucus of brook charr (Salvelinus fontinalis) against two major pathogens Flavobacterium columnare and Flavobacterium psychrophilum. Nine bacterial strains were isolated from brook charr skin mucus and tested for potential antagonistic activity. Results from both agar diffusion assays and broth co-culture assays showed the presence of antagonism. We identified seven bacterial strains, collected from unstressed fish, which exerted strong antagonism against F. psychrophilum and/or F. columnare. These strains were mixed and used to treat columnaris disease in an in vivo experiment in which four distinct fish families were tested. This treatment resulted in a decrease of mortality (54-86%) across fish families indicating that candidates from the host microbiota are potentially suitable for probiotic development. This would allow for the efficient (ability to adhere and colonize the host mucus) and durable management (antagonistic effect against pathogens which would be harmless for the host and safe for its environment) of opportunistic diseases in aquaculture.

Fast Transcriptional Responses to Domestication in the Brook Charr Salvelinus fontinalis

Domestication has been practiced for centuries yet directed toward relatively few terrestrial crops and animals. While phenotypic and quantitative genetic changes associated with domestication have been amply documented, little is known about the molecular changes underlying the phenotypic evolution during the process. Here, we have investigated the brook charr (Salvelinus fontinalis) responses to artificial selection by means of transcriptional analysis of ∼32,000 cDNA features performed in both selected and control populations reared under identical environmental conditions during four generations. Our results indicate that selective breeding led to significant changes in the transcription of genes at the juvenile stage, where we observed 4.16% (156/3750) of differentially expressed genes between the two lines. No significant genes were revealed at the earlier life stage. Moreover, when comparing our results to those of previous studies on Atlantic salmon that compared lines that were selected for five to seven generations for similar traits (e.g., growth), genes with similar biological functions were found to be under selection in both studies. These observations indicate that (1) four generations of selection caused substantial changes in regulation of gene transcription between selected and control populations and (2) selective breeding for improving the same phenotypic traits (e.g., rapid growth) in brook charr and Atlantic salmon tended to select for the same changes in transcription profiles as the expression of a small and similar set of genes was affected by selection.

Major disruption of gene expression in hybrids between young sympatric anadromous and resident populations of brook charr (Salvelinus fontinalis Mitchill)

Genome‐wide analyses of the transcriptome have suggested that male‐biased genes are the first targets of genomic incompatibilities (g.i.) in inter‐specific hybrids. However, those studies have almost invariably focused on Drosophila species that diverged at least 0.9 Ma, and with sterile male hybrids. Here, we use microarrays to analyse patterns of gene expression in very closely related (divergence <12 000 years), sympatric, but ecologically divergent anadromous and resident populations of brook charr (Salvelinus fontinalis) and their F1 hybrids. Our results show a dramatic breakdown of gene expression patterns in hybrids compared with their parental relatives. Several disrupted genes are related to energetic metabolism, immune response, osmoregulation and protection against oxidative stress, and none has sex‐biased functions. Besides, pure individuals show no expression differences at most of the genes disrupted in hybrids, which may suggest the operation of some form of stabilizing selection. Taken together, these results both confirm the idea that perturbations of regulatory networks represent a significant source of g.i. and support the suggestion that developmental pathways can diverge through time without any manifest change in the phenotypic outcome. While the role of other evolutionary forces (e.g. genetic drift) cannot be ruled out, this study suggests that ecological selective processes may provide the initial driving force behind disruption of gene expression in inter‐specific hybrids.