Circulating levels of plasma IGF-I during recovery from size-selective harvesting in Menidia menidia.

Selection for growth-related traits in domesticated fishes often results in predictable changes within the growth hormone-insulin-like growth factor (GH-IGF-1) axis. Little is known about the mechanisms controlling changes in growth capacity resulting from fishery-induced evolution. We took advantage of a long-term study where Menidia menidia were selected for size at age over multiple generations to mimic fisheries-induced selection. This selection regime produced three populations with significant differences in intrinsic growth rate. These growth differences partially rebounded, but persisted even after selection was relaxed, resulting in fast, intermediate, and slow-growing lines. Plasma IGF-1 was measured in these populations as a potential target of selection on growth. IGF-1 was significantly correlated with current length and mass, and was positively correlated with growth rate (g d(-1)) in two lines, indicating it may be an appropriate indicator of growth capacity. The slow-growing line exhibited higher overall IGF-1 levels relative to the depressed IGF-1 seen in the fast-growing line, contrary to our prediction. We offer possible explanations for this unusual pattern and argue that somatic growth is likely to be under control of mechanism(s) downstream to IGF-1. IGF-1 provides an interesting basis for understanding endocrine control of growth in response to artificial selection and recovery.

Spatial scale and divergent patterns of variation in adapted traits in the ocean.

The geography of adaptive genetic variation is crucial to species conservation yet poorly understood in marine systems. We analyse the spatial scale of genetic variation in traits that broadly display adaptation throughout the range of a highly dispersive marine species. We conducted common garden experiments on the Atlantic silverside, Menidia menidia, from 39 locations along its 3000 km range thereby mapping genetic variation for growth rate, vertebral number and sex determination. Each trait displayed unique clinal patterns, with significant differences (adaptive or not) occurring over very small distances. Breakpoints in the cline differed among traits, corresponding only partially with presumed eco-geographical boundaries. Because clinal patterns are unique to each selected character, neutral genes or those coding for a single character cannot serve as proxies for the genetic structure as a whole. Conservation plans designed to protect essential genetic subunits of a species will need to account for such complex spatial structures.

Adaptation to climate change: contrasting patterns of thermal-reaction-norm evolution in Pacific versus Atlantic silversides.

How organisms may adapt to rising global temperatures is uncertain, but concepts can emerge from studying adaptive physiological trait variations across existing spatial climate gradients. Many ectotherms, particularly fish, have evolved increasing genetic growth capacities with latitude (i.e. countergradient variation (CnGV) in growth), which are thought to be an adaptation primarily to strong gradients in seasonality. In contrast, evolutionary responses to gradients in mean temperature are often assumed to involve an alternative mode, 'thermal adaptation'. We measured thermal growth reaction norms in Pacific silverside populations (Atherinops affinis) occurring across a weak latitudinal temperature gradient with invariant seasonality along the North American Pacific coast. Instead of thermal adaptation, we found novel evidence for CnGV in growth, suggesting that CnGV is a ubiquitous mode of reaction-norm evolution in ectotherms even in response to weak spatial and, by inference, temporal climate gradients. A novel, large-scale comparison between ecologically equivalent Pacific versus Atlantic silversides (Menidia menidia) revealed how closely growth CnGV patterns reflect their respective climate gradients. While steep growth reaction norms and increasing growth plasticity with latitude in M. menidia mimicked the strong, highly seasonal Atlantic coastal gradient, shallow reaction norms and much smaller, latitude-independent growth plasticity in A. affinis resembled the weak Pacific latitudinal temperature gradient.

High connectivity among locally adapted populations of a marine fish (Menidia menidia).

Patterns of connectivity are important in understanding the geographic scale of local adaptation in marine populations. While natural selection can lead to local adaptation, high connectivity can diminish the potential for such adaptation to occur. Connectivity, defined as the exchange of individuals among subpopulations, is presumed to be significant in most marine species due to life histories that include widely dispersive stages. However, evidence of local adaptation in marine species, such the Atlantic silverside, Menidia menidia, raises questions concerning the degree of connectivity. We examined geochemical signatures in the otoliths, or ear bones, of adult Atlantic silversides collected in 11 locations along the northeastern coast of the United States from New Jersey to Maine in 2004 and eight locations in 2005 using laser ablation inductively coupled plasma mass spectrometry (ICP-MS) and isotope ratio monitoring mass spectrometry (irm-MS). These signatures were then compared to baseline signatures of juvenile fish of known origin to determine natal origin of these adult fish. We then estimated migration distances and the degree of mixing from these data. In both years, fish generally had the highest probability of originating from the same location in which they were captured (0.01-0.80), but evidence of mixing throughout the sample area was present. Furthermore, adult M. menidia exhibit highly dispersive behavior with some fish migrating over 700 km. The probability of adult fish returning to natal areas differed between years, with the probability being, on average, 0.2 higher in the second year. These findings demonstrate that marine species with largely open populations are capable of local adaptation despite apparently high gene flow.

Footprints of local adaptation span hundreds of linked genes in the Atlantic silverside genome.

The study of local adaptation in the presence of ongoing gene flow is the study of natural selection in action, revealing the functional genetic diversity most relevant to contemporary pressures. In addition to individual genes, genome-wide architecture can itself evolve to enable adaptation. Distributed across a steep thermal gradient along the east coast of North America, Atlantic silversides (Menidia menidia) exhibit an extraordinary degree of local adaptation in a suite of traits, and the capacity for rapid adaptation from standing genetic variation, but we know little about the patterns of genomic variation across the species range that enable this remarkable adaptability. Here, we use low-coverage, whole-transcriptome sequencing of Atlantic silversides sampled along an environmental cline to show marked signatures of divergent selection across a gradient of neutral differentiation. Atlantic silversides sampled across 1371 km of the southern section of its distribution have very low genome-wide differentiation (median F ST = 0.006 across 1.9 million variants), consistent with historical connectivity and observations of recent migrants. Yet almost 14,000 single nucleotide polymorphisms (SNPs) are nearly fixed (F ST > 0.95) for alternate alleles. Highly differentiated SNPs cluster into four tight linkage disequilibrium (LD) blocks that span hundreds of genes and several megabases. Variants in these LD blocks are disproportionately nonsynonymous and concentrated in genes enriched for multiple functions related to known adaptations in silversides, including variation in lipid storage, metabolic rate, and spawning behavior. Elevated levels of absolute divergence and demographic modeling suggest selection maintaining divergence across these blocks under gene flow. These findings represent an extreme case of heterogeneity in levels of differentiation across the genome, and highlight how gene flow shapes genomic architecture in continuous populations. Locally adapted alleles may be common features of populations distributed along environmental gradients, and will likely be key to conserving variation to enable future responses to environmental change.

Reversal of evolutionary downsizing caused by selective harvest of large fish.

Evolutionary responses to the long-term exploitation of individuals from a population may include reduced growth rate, age at maturation, body size and productivity. Theoretical models suggest that these genetic changes may be slow or impossible to reverse but rigorous empirical evidence is lacking. Here, we provide the first empirical demonstration of a genetically based reversal of fishing-induced evolution. We subjected six populations of silverside fish (Menidia menidia) to three forms of size-selective fishing for five generations, thereby generating twofold differences among populations in mean weight and yield (biomass) at harvest. This was followed by an additional five generations during which size-selective harvest was halted. We found that evolutionary changes were reversible. Populations evolving smaller body size when subjected to size-selective fishing displayed a slow but significant increase in size when fishing ceased. Neither phenotypic variance in size nor juvenile survival was reduced by the initial period of selective fishing, suggesting that sufficient genetic variation remained to allow recovery. By linear extrapolation, we predict full recovery in about 12 generations, although the rate of recovery may taper off near convergence. The recovery rate in any given wild population will also depend on other agents of selection determined by the specifics of life history and environment. By contrast, populations that in the first five generations evolved larger size and yield showed little evidence of reversal. These results show that populations have an intrinsic capacity to recover genetically from harmful evolutionary changes caused by fishing, even without extrinsic factors that reverse the selection gradient. However, harvested species typically have generation times of 3-7 years, so recovery may take decades. Hence, the need to account for evolution in managing fisheries remains.

Contrasting genomic shifts underlie parallel phenotypic evolution in response to fishing.

Humans cause widespread evolutionary change in nature, but we still know little about the genomic basis of rapid adaptation in the Anthropocene. We tracked genomic changes across all protein-coding genes in experimental fish populations that evolved pronounced shifts in growth rates due to size-selective harvest over only four generations. Comparisons of replicate lines show parallel allele frequency shifts that recapitulate responses to size-selection gradients in the wild across hundreds of unlinked variants concentrated in growth-related genes. However, a supercluster of genes also rose rapidly in frequency and dominated the evolutionary dynamic in one replicate line but not in others. Parallel phenotypic changes thus masked highly divergent genomic responses to selection, illustrating how contingent rapid adaptation can be in the face of strong human-induced selection.

Mercury bioaccumulation increases with latitude in a coastal marine fish (Atlantic silverside, Menidia menidia).

Human exposure to the neurotoxic methylmercury (MeHg) occurs primarily via the consumption of marine fish, but the processes underlying large-scale spatial variations in fish MeHg concentrations [MeHg], which influence human exposure, are not sufficiently understood. We used the Atlantic silverside (Menidia menidia), an extensively studied model species and important forage fish, to examine latitudinal patterns in total Hg [Hg] and [MeHg]. Both [Hg] and [MeHg] significantly increased with latitude (0.014 and 0.048 µg MeHg g-1 dw per degree of latitude in juveniles and adults, respectively). Four known latitudinal trends in silverside traits help explain these patterns: latitudinal increase in MeHg assimilation efficiency, latitudinal decrease in MeHg efflux, latitudinal increase in weight loss due to longer and more severe winters, and latitudinal increase in food consumption as an adaptation to decreasing length of the growing season. Given the absence of a latitudinal pattern in particulate MeHg, a diet proxy for zooplanktivorous fish, we conclude that large-scale spatial variation in growth is the primary control of Hg bioaccumulation in this and potentially other fish species.

Maladaptive changes in multiple traits caused by fishing: impediments to population recovery.

Some overharvested fish populations fail to recover even after considerable reductions in fishing pressure. The reasons are unclear but may involve genetic changes in life history traits that are detrimental to population growth when natural environmental factors prevail. We empirically modelled this process by subjecting populations of a harvested marine fish, the Atlantic silverside, to experimental size-biased fishing regimes over five generations and then measured correlated responses across multiple traits. Populations where large fish were selectively harvested (as in most fisheries) displayed substantial declines in fecundity, egg volume, larval size at hatch, larval viability, larval growth rates, food consumption rate and conversion efficiency, vertebral number, and willingness to forage. These genetically based changes in numerous traits generally reduce the capacity for population recovery.

Evolution of intrinsic growth rate: metabolic costs drive trade-offs between growth and swimming performance in Menidia menidia.

There is strong evidence that genetic capacity for growth evolves toward an optimum rather than an absolute maximum. This implies that fast growth has a cost and that trade-offs occur between growth and other life-history traits, but the fundamental mechanisms are poorly understood. Previous work on the Atlantic silverside fish Menidia menidia has demonstrated a trade-off between growth and swimming performance. We hypothesize that the trade-off derives from the competing metabolic demands associated with growth and swimming activity. We tested this by measuring standard metabolic rate (M(STD)), maximum sustainable metabolic rate (M(ACT)) and metabolic scope of laboratory-reared silversides originating from two geographically distinct populations with well-documented differences in genetic capacity for growth. The fast-growth genotype had a significantly greater M(STD) than the slow-growth genotype, but a similar MACT when swum to near exhaustion. The scope for activity of the fast-growth genotype was lower than that of the slow-growth genotype. Furthermore, the fast-growth genotype eats larger meals, thereby incurring a greater postprandial oxygen demand. We conclude that a metabolic trade-off occurs between growth and other metabolic demands and that this trade-off provides a general mechanism underlying the evolution of growth rate.

Tests of bioaccumulation models for polychlorinated biphenyl compounds: a study of young-of-the-year bluefish in the Hudson River estuary, USA.

A field-based study regarding uptake of polychlorinated biphenyl compounds (PCBs) by young-of-the-year (YOY) bluefish (Pomatomus saltatrix) was initiated to test a steady-state model of bioaccumulation and trophic transfer in a rapidly growing fish. Determination of prey composition as well as size-dependent growth and specific consumption rates for YOY bluefish from separate field and laboratory studies enabled the input of these species-specific parameters into the model. Furthermore, the time and duration of the exposure of YOY bluefish to dissolved PCBs from a well-characterized system (Hudson River, USA) was well known. Patterns of accumulation of individual PCB congeners differed relative to the accumulation of total PCBs, with the greatest net accumulation occurring for the higher-molecular-weight congeners. Comparison of lipid-normalized bioaccumulation factors (BAFs) with the octanol-water partition coefficients of individual PCB congeners revealed bluefish to be above the BAFs predicted by lipid-based equilibrium partitioning, suggesting that uptake from food is an important source of PCBs in YOY bluefish. Comparison of measured BAFs with values predicted by a steady-state, food-chain model showed good first-order agreement.

Pattern and scale of geographic variation in environmental sex determination in the Atlantic silverside, Menidia menidia.

The Atlantic silverside, Menidia menidia (Pisces: Atherinidae), exhibits an exceptionally high level of clinal variation in sex determination across its geographic range. Previous work suggested linear changes in the level of temperature-dependent sex determination (TSD) with increasing latitude. Based on comparisons at 31 sites encompassing the entire species' range, we find that the change in level of TSD with latitude is instead highly nonlinear. The level of TSD is uniformly high in the south (Florida to New Jersey), then declines rapidly into the northern Gulf of Maine where genotypic sex determination (GSD) predominates and then rebounds to moderate levels of TSD in the northern-most populations of the Gulf of St. Lawrence. Major latitudinal breakpoints occur in central New Jersey (40(o)N) and the northern Gulf of Maine (44(o)N). No populations display pure TSD or GSD. Length of the growing season is the likely agent of selection driving variation in TSD with a threshold at 210 days. Because gene flow among populations is high, such distinct patterns of geographic variation in TSD/GSD are likely maintained by contemporary selection thereby demonstrating the adaptive fine tuning of sex determining mechanisms.

Rapid growth results in increased susceptibility to predation in Menidia menidia.

Several recent studies have demonstrated that rapid growth early in life leads to decreased physiological performance. Nearly all involved experiments over short time periods (<1 day) to control for potentially confounding effects of size. This approach, however, neglects the benefits an individual accrues by growing. The net effect of growth can only be evaluated over a longer interval in which rapidly growing individuals are allowed the time required to attain the expected benefits of large size. We used two populations of Menidia menidia with disparate intrinsic growth rates to address this issue. We compared growth and survivorship among populations subject to predation in mesocosms under ambient light and temperature conditions for a period of up to 30 days to address two questions: Do the growth rates of fish in these populations respond differently to the presence of predators? Is the previously demonstrated survival cost of growth counterbalanced by the benefits of increased size? We found that growth was insensitive to predation risk: neither population appeared to modify growth rates in response to predation levels. Moreover, the fast-growing population suffered significantly higher mortality throughout the trials despite being 40% larger than the slow-growing population at the experiment's end. These results confirm that the costs of rapid growth extend over prolonged intervals and are not ameliorated merely by the attainment of large size.

Nonlinear growth cost in Menidia menidia: theory and empirical evidence.

Juvenile growth is submaximal in many species, suggesting that a trade-off with juvenile growth must exist. In support of this, recent studies have demonstrated that rapid growth early in life results in decreased physiological performance. Theory clearly shows that for submaximal growth in juveniles to be optimal, the cost of growth must be nonlinear. However, nearly all of the empirical evidence for costs of growth comes from linear comparisons between fast- and slow-growing groups. It is consequently unclear whether any known cost can account for the evolution of submaximal juvenile growth. To test whether the cost of growth exhibits the logically necessary nonlinearity, we measured critical swimming speed (Ucrit), the maximum speed sustained in incremental velocity trials, in Atlantic silversides, a species for which the costs and benefits of growth are well studied. To increase our ability to detect a nonlinear relationship between Ucrit, a proxy for juvenile fitness, and growth, we manipulated ration levels to produce a broad range of growth rates (0.16 mm/day(-1) to 1.20 mm/day(-1)). Controlling for size and age, we found that Ucrit decreased precipitously as growth approached the physiological maximum. Using Akaike's information criterion, we show that swimming performance decreases with the square of growth rate, providing the first demonstration of a nonlinear cost of growth.

Latitudinal differences in somatic energy storage: adaptive responses to seasonality in an estuarine fish (Atherinidae: Menidia menidia).

This study focuses on the seasonal accumulation and depletion of somatic energy in the Atlantic silverside (Menidia menidia), an annual estuarine fish. Previous research revealed that northern silversides are subject to strong size-dependent winter mortality, while southern fish suffer no appreciable winter mortality. To examine whether there was geographic differentiation in allocation strategies, we compared temporal patterns of energy storage and utilization among three populations along this gradient in seasonality. The comparative design used monthly or biweekly samples of fish collected in the wild, as well as samples of fish from each population reared in a common environment, where genetic differences can be clarified. Somatic energy stores were quantified via gravimetric analysis of neutral storage lipids and lean tissue. Analysis revealed that small individuals maintained relatively low levels of lipid reserves, which may account for their lower survival in winter. Wild fish in the north rapidly accumulated large somatic reserves, which were depleted over the winter and then increased again during the subsequent spring breeding season. In wild southern fish, relatively small reserves accumulated slowly until breeding commenced in the spring. The common-environment comparison of somatic storage patterns revealed a genetic basis for among-population differences in reserve accumulation rates, but no differences in the amount of reserves stored. We conclude that the overwinter depletion of somatic reserves has a significant selective impact on energy accumulation and allocation strategies in seasonal environments.

The allometry of energy reserve depletion: test of a mechanism for size-dependent winter mortality.

We experimentally tested the hypothesis that energy reserve depletion varies inversely with size in the fish Menidia menidia, an estuarine fish known to exhibit size-dependent winter mortality. Individuals in two size groups were starved at two winter temperatures (4°and 8°C) and sacrificed at a range of time intervals (up to 127 days). Lipid levels and lean tissue were analyzed to estimate somatic energy storage. As predicted, energy depletion was greater at high temperatures, and proportionally greater in small than in large fish. After 60 days of starvation at 4°C, small fish retained an average of 67% of their original energy reserves (vs 53% at 8°C), while large fish retained an average of 80% (vs 66% at 8°C). At 4°C, fish that were fed depleted their energy reserves as rapidly as unfed fish, but at 8°C, fish that were fed maintained reserves at higher levels than unfed fish. A high proportion of unfed fish (56% at 4°C, 27% at 8°C) died before they were to be sacrificed. Survival probability did not vary with size, nor was it influenced by the amount of energy reserves. The rate of energy depletion (equivalent to routine metabolic rate) decreased gradually over time, particularly in small fish. Routine metabolism did not conform to a single scaling relationship. Within each temperature-size group, the routine rate declined more rapidly than metabolically active mass (lean mass). At 8°C, the difference between size groups in energy depletion rate conformed closely to the expected allometry exponent of 0.8. In contrast, at 4°C, the estimated allometry exponent increased over the experiment (-0.19 to 2.5). We conclude that strategies to minimize energy loss may often modify bioenergetic scaling relationships.

Countergradient variation in growth rate: compensation for length of the growing season among Atlantic silversides from different latitudes.

How do organisms adapt to the differences in temperature and length of the growing season that occur with latitude? Among Atlantic silversides (Menidia menidia) along the east coast of North America, the length of the first growing season declines by a factor of about 2.5 with increasing latitude. Yet body size at the end of the first growing season does not decline. High-latitude fish must, therefore, grow faster within the growing season than do low-latitude fish. This geographical pattern has a genetic basis. Laboratory experiments on fish from six different locations revealed a latitudinal gradient in the capacity for growth (i.e., maximum growth potential). In two subsequent experiments using fish from Nova Scotia (NS), New York (NY) and South Carolina (SC) that had been separately reared in a common environment for several generations, differences in growth rate among populations were highly significant. The rank order was NS>NY>SC, but the difference among populations depended on temperature. High-latitude fish outperformed those from low latitudes primarily at the high temperatures that low-latitude fish would be expected to experience most often in nature. These results suggest that instead of being adapted for growth at low temperatures, fish from high latitudes are adapted for rapid elevation of growth rate during the brief interval of the year when high temperatures occur. Selection on growth rate results from sizedependent winter mortality: the importance to winter survival of being large increases with latitude but the length of the growing season simultaneously decreases. The end result is countergradient variation in growth rate, a phenomenon that may be much more widespread than currently recognized.

The covariance of routine and compensatory juvenile growth rates over a seasonality gradient in a coastal fish.

Compensatory growth is widespread in juvenile animals; it refers to the ability to grow at faster-than-routine rates, following intervals of slow growth due to reduced food supply or temperature. Whether the ability to grow rapidly under routine conditions is associated with enhanced or reduced compensatory growth performance is unknown. We examine the among-population covariance in routine and compensatory growth ability in juvenile Atlantic silverside, Menidia menidia. Routine growth rate in this species positively varies with latitude, indicating selection for more rapid increase in size in highly seasonal environments. High-latitude fish may also show strong compensatory ability, considering the selective value of rapid recovery from winter for growth and breeding in the spring. Alternatively, low-latitude fish may be more readily able to compensate because they have more energetic scope for growth. To test these alternatives, laboratory-reared juveniles from three populations [Nova Scotia (NS), New York (NY), and South Carolina (SC)] were maintained at temperatures promoting maximum growth. In each population, control fish were furnished ad libitum rations, while treatment fish were subject to 5 or 10 days of maintenance ration before returning to unlimited ration. Treatment fish grew slightly in length, but not mass, over the period of limited ration. We observed compensatory growth in all populations. NS fish recovering from 10 days of limited ration grew 12% faster in length and 46% faster in dry mass than NS control fish, over 15 days of unrestricted growth. In contrast, recovering SC fish grew 1.4% faster in length and 22% faster in dry mass than SC control fish. A period of starvation (mass loss) is not a required condition for a compensatory response, as has been assumed in theoretical work. The positive covariance between routine and compensatory growth performance indicates that the growth rate strategy of high latitude fish includes rapid recovery from periods of depletion, adding to the suite of characteristics that have differentiated along an environmental gradient in seasonality.