Estuarine Fish Fundulus Heteroclitus Research Articles

Stable genetic structure andconnectivity in pollution-adapted and nearby pollution-sensitive populations of Fundulus heteroclitus.

Populations of the non-migratory estuarine fish Fundulus heteroclitus inhabiting the heavily polluted New Bedford Harbour (NBH) estuary have shown inherited tolerance to local pollutants introduced to their habitats in the past 100 years. Here we examine two questions: (i) Is there pollution-driven selection on the mitochondrial genome across a fine geographical scale? and (ii) What is the pattern of migration among sites spanning a strong pollution gradient? Whole mitochondrial genomes were analysed for 133 F. heteroclitus from seven nearby collection sites: four sites along the NBH pollution cline (approx. 5 km distance), which had pollution-adapted fish, as well as one site adjacent to the pollution cline and two relatively unpolluted sites about 30 km away, which had pollution-sensitive fish. Additionally, we used microsatellite analyses to quantify genetic variation over three F. heteroclitus generations in both pollution-adapted and sensitive individuals collected from two sites at two different time points (1999/2000 and 2007/2008). Our results show no evidence for a selective sweep of mtDNA in the polluted sites. Moreover, mtDNA analyses revealed that both pollution-adapted and sensitive populations harbour similar levels of genetic diversity. We observed a high level of non-synonymous mutations in the most polluted site. This is probably associated with a reduction in Ne and concomitant weakening of purifying selection, a demographic expansion following a pollution-related bottleneck or increased mutation rates. Our demographic analyses suggest that isolation by distance influences the distribution of mtDNA genetic variation between the pollution cline and the clean populations at broad spatial scales. At finer scales, population structure is patchy, and neither spatial distance, pollution concentration or pollution tolerance is a good predictor of mtDNA variation. Lastly, microsatellite analyses revealed stable population structure over the last decade.

Growth and Movements of Mummichogs (Fundulus heteroclitus) Along Armored and Vegetated Estuarine Shorelines

Alteration of estuarine shorelines associated with increased urbanization can significantly impact biota and food webs. This study determined the impact of shoreline alteration on growth and movement of the estuarine fish Fundulus heteroclitus in a tributary of the Delaware Coastal Bays. Fundulus heteroclitus is abundant along the east coast of the USA, and is an important trophic link between marsh and subtidal estuary. The restricted home range of F. heteroclitus allowed discrete sampling, and fish growth comparisons, along 35–65-m long stretches of fringing Spartina alterniflora and Phragmites australis marsh, riprap, and bulkhead. Fundulus heteroclitus were tagged with decimal Coded Wire Tags. Of 725 tagged F. heteroclitus, 89 were recaptured 30–63 days later. Mean growth rate (0.06–0.15 mm day−1 across all shoreline types) was greatest at riprap, lowest at Spartina and Phragmites, and intermediate at bulkhead, where growth was not significantly different from any other shoreline. This suggests that discernible environments exist along different shoreline types, even at the scale of tens of meters. No difference in movement distance was detected at different shoreline types; most individuals displayed a high degree of site fidelity. Forty-seven percent were recaptured within 5 m of their tagging location, although alongshore movements up to 475 m were recorded. Estimates of relative F. heteroclitus productivity, using relative density data from a concurrent study, were highest along Spartina and Phragmites, intermediate at riprap, and lowest at bulkhead. Therefore, despite greater growth rates along riprap than at vegetated shores, armoring reduces abundance sufficiently to negatively impact localized productivity of F. heteroclitus.

Sequence and functional characterization of hypoxia-inducible factors, HIF1α, HIF2αa, and HIF3α, from the estuarine fish, Fundulus heteroclitus.

The hypoxia-inducible factor (HIF) family of transcription factors plays central roles in the development, physiology, pathology, and environmental adaptation of animals. Because many aquatic habitats are characterized by episodes of low dissolved oxygen, fish represent ideal models to study the roles of HIF in the response to aquatic hypoxia. The estuarine fish Fundulus heteroclitus is found in habitats prone to hypoxia. It responds to low oxygen via behavioral, physiological, and molecular changes, and one member of the HIF family, HIF2α, has been previously described. Herein, cDNA sequencing, phylogenetic analyses, and genomic approaches were used to determine other members of the HIFα family from F. heteroclitus and their relationships to HIFα subunits from other vertebrates. In vitro and cellular approaches demonstrated that full-length forms of HIF1α, HIF2α, and HIF3α independently formed complexes with the β-subunit, aryl hydrocarbon receptor nuclear translocator, to bind to hypoxia response elements and activate reporter gene expression. Quantitative PCR showed that HIFα mRNA abundance varied among organs of normoxic fish in an isoform-specific fashion. Analysis of the F. heteroclitus genome revealed a locus encoding a second HIF2α-HIF2αb-a predicted protein lacking oxygen sensing and transactivation domains. Finally, sequence analyses demonstrated polymorphism in the coding sequence of each F. heteroclitus HIFα subunit, suggesting that genetic variation in these transcription factors may play a role in the variation in hypoxia responses among individuals or populations.

Evolution of tolerance to PCBs and susceptibility to a bacterial pathogen ( Vibrio harveyi) in Atlantic killifish ( Fundulus heteroclitus) from New Bedford (MA, USA) harbor

A population of the non-migratory estuarine fish Fundulus heteroclitus (Atlantic killifish) resident to New Bedford (NB), Massachusetts, USA, an urban harbor highly contaminated with polychlorinated biphenyls (PCBs), demonstrates recently evolved tolerance to some aspects of PCB toxicity. PCB toxicology, ecological theory, and some precedence supported expectations of increased susceptibility to pathogens in NB killifish. However, laboratory bacterial challenges of the marine pathogen Vibrio harveyi to wild fish throughout the reproductive season and to their mature laboratory-raised progeny demonstrated comparable survival by NB and reference killifish, and improved survival by NB males. These results are inconsistent with hypothesized trade-offs of adaptation, and suggest that evolved tolerance in NB killifish may include mechanisms that minimize the immunosuppressive effects of PCBs. Compensatory strategies of populations persisting in highly contaminated environments provide a unique perspective for understanding the long-term ecological effects of toxic chemicals.

Population genetic structure of a nonmigratory estuarine fish (Fundulus heteroclitus) across a strong gradient of polychlorinated biphenyl contamination

Populations of the estuarine fish Fundulus heteroclitus indigenous to contaminated sites exhibit heritable resistance to some of the toxic effects of early life-stage exposure to polychlorinated biphenyls (PCBs). This evolved tolerance provides evidence of strong selection by PCBs, and it suggests other potential genetic effects of these stressors on resident populations. Environmental contaminants have the potential to affect the genetic structure of populations and to reduce genetic diversity, but species life-history traits, particularly patterns of migration and dispersal, also influence the distribution of genetic variation among populations. Therefore, the present was conducted to determine whether genetic diversity or genetic structure is altered in populations of F. heteroclitus indigenous to 18 sites in Massachusetts (USA) and Rhode Island (USA), representing a steep gradient of sediment PCB contamination and culminating in a Superfund site at New Bedford Harbor (NBH; MA, USA). Allele frequencies at enzymatic loci were used to assess genetic structure and diversity. Selection experiments using a highly toxic PCB congener (3,3',4,4',5-pentachlorobiphenyl) were conducted to determine if genetic patterns at field sites could be associated with contaminant exposures. Although allele frequencies clearly reflected a pattern of isolation by distance, the results indicated neither significant loss of genetic diversity nor alteration of allele frequencies for populations of F. heteroclitus in NBH.

Strong positive selection and habitat-specific amino acid substitution patterns in MHC from an estuarine fish under intense pollution stress.

Population-level studies using the major histocompatibility complex (Mhc) have linked specific alleles with specific diseases, but data requirements are high and the power to detect disease association is low. A novel use of Mhc population surveys involves mapping allelic substitutions onto the inferred structural molecular model to show functional differentiation related to local selective pressures. In the estuarine fish Fundulus heteroclitus, populations experiencing strong differences in antigenic challenges show significant differences in amino acid substitution patterns that are reflected as variation in the structural location of changes between populations. Fish from a population genetically adapted to severe chemical pollution also show novel patterns of DNA substitution at a highly variable Mhc class II B locus including strong signals of positive selection at inferred antigen-binding sites and population-specific signatures of amino acid substitution. Heavily parasitized fish from an extreme PCB-contaminated (U.S. Environmental Protection Agency Superfund) site show enhanced population-specific substitutions in the a-helix portion of the inferred antigen-binding region. In contrast, fish from an unpolluted site show a significantly different pattern focused on the first strand of the B-pleated sheet. Whether Mhc population profile differences represent the direct effects of chemical toxicants or indirect parasite-mediated selection, the result is a composite habitat-specific signature of strong selection and evolution affecting the genetic repertoire of the major histocompatibility complex.

Effects of benzo[ a]pyrene exposure on a fish population resistant to the toxic effects of dioxin-like compounds

Effects of a model polycyclic aromatic hydrocarbon (PAH) were compared in populations of the estuarine fish Fundulus heteroclitus indigenous to a reference site and one highly contaminated with polychlorinated biphenyls (PCBs) and other compounds. The fish population resident to the PCB-contaminated site is genetically resistant to those PCB congeners categorized as dioxin-like compounds (DLCs) that act through the aryl hydrocarbon receptor (AHR). In response to DLC exposures, these DLC-resistant fish showed poor inducibility for enzymes known to be regulated by the AHR pathway and important for the metabolism of xenobiotics including some PAHs that also act as AHR agonists. Therefore, a laboratory study using the model PAH, benzo[ a]pyrene (BaP), was conducted to evaluate how PAHs might affect these wild fish populations that differed in their inherent sensitivities to DLCs and in their tissue concentrations of contaminants. Following BaP treatment, the activities of two xenobiotic metabolizing enzymes and the concentrations of BaP-DNA adducts, as measured using the 32P-postlabeling method, were lower in the livers of DLC-resistant than reference fish. These results suggest that DLC-resistance could provide protection following chronic exposures to PAHs from the long-term consequences of DNA adduct formation, such as cancer. Alternatively, reduced metabolism and elimination of toxic or photo-activated PAHs could have acute consequences to the health and reproduction of these DLC-resistant fish and their progeny. These fish populations provide useful models to evaluate the potential costs and benefits of genetic adaptation in wildlife populations subject to anthropogenic stress.

Stored retinoids in populations of the estuarine fish Fundulus heteroclitus indigenous to PCB-contaminated and reference sites.

Concentrations of retinoids, derivatives of vitamin A, were measured in populations of the nonmigratory estuarine fish Fundulus heteroclitus, indigenous to a reference site and a site highly contaminated with polychlorinated biphenyls (PCBs) to address the hypothesis that contaminant exposure causes depletion of stored retinoids in native fish populations. To assess differences related to chemical exposure, as well as season and diet, hepatic retinoid analyses were conducted using fish collected early and late in the natural spawning season and after laboratory holding. Though hepatic retinoid composition was generally similar among groups, hepatic retinoid concentrations differed. Laboratory-held fish had higher hepatic retinyl ester concentrations than field-collected fish. Among field-collected fish, those collected early in the spawning season had higher hepatic retinyl ester concentrations than those collected later in the spawning season. Although there was no evidence of the dramatic retinoid depletions that have been reported in highly exposed populations of other fish species, hepatic retinoid stores were consistently lower in F. heteroclitus indigenous to the highly PCB-contaminated site. These results are consistent with prior findings that fish from this contaminated site are relatively insensitive to some of the toxic effects of PCB exposures, including retinoid depletion.

Adaptations of wild populations of the estuarine fish Fundulus heteroclitus to persistent environmental contaminants

Many aquatic species, including the estuarine fish Fundulus heteroclitus (mummichogs), adapt to local environmental conditions. We conducted studies to evaluate whether highly exposed populations of mummichogs adapt to toxic environmental contaminants. These fish populations are indigenous to an urban estuary contaminated with persistent and bioaccumulative contaminants (dioxin-like compounds, or DLCs) that are particularly toxic to the early development of fish. We conducted laboratory challenge experiments to compare mummichog embryos and larvae from reference sites and this highly contaminated site [New Bedford Harbor (NBH), Massachusetts, USA] for their sensitivity to DLCs. While there was variation in DLC-responsiveness within each group, fish from NBH were profoundly less sensitive to DLCs than reference fish. Specifically, concentrations of DLCs similar to those measured in NBH-collected mummichog eggs were lethal to reference embryos. Further, DLC-responsiveness was inherited and independent of maternal contaminant contributions. These findings are consistent with the conclusion that DLC contamination in NBH has contributed to the selection of fish that are resistant to the short-term toxic effects of these environmental-contaminant exposures. This adaptation may be a critical mechanism by which fish populations persist in this highly contaminated site. Further evaluation of this ecosystem may provide important information concerning the direct and indirect consequences of this “unnatural” selection.

Molecular cloning of CYP1A from the estuarine fish Fundulus heteroclitus and phylogenetic analysis of CYP1 genes: update with new sequences

Since we published a phylogenetic analysis of the CYP1A subfamily in 1995, several additional full-length sequences have been reported, including three members of an entirely new subfamily, CYP1B. Two avian sequences were recently published, so that CYP1A sequence data are now available from three of the five major vertebrate lineages. The two new branches that have been added to the CYP1 family tree significantly add to our understanding of P450 evolution. The inclusion of the CYP1Bs to the phylogenetic analysis allows us to root inferred trees. Addition of the avian CYP1As indicates that the CYP1A1/CYP1A2 duplication present in the mammalian lineage may have occurred after the divergence of birds and mammals. The number of fish species from which full-length coding regions of CYP1A genes have been sequenced has increased from four (trout, plaice, toadfish, and scup) to nine. These include CYP1A sequences from tomcod, butterflyfish, sea bream, sea bass, and the full-length sequence of CYP1A from the killifish Fundulus heteroclitus that is reported here. Phylogenetic analyses incorporating the new fish CYP1A sequences support our original conclusion that the fish CYP1As are monophyletic and indicate that the genes are evolving at very different rates in different species.