Transgenerational effects of parental crude oil exposure on the morphology of adult Fundulus grandis
Introduction
The 2010 Deepwater Horizon oil spill resulted in the release of an estimated 507 million liters of crude oil into the northern Gulf of Mexico (nGoM) and oiled 2100 km of coastline (Deepwater Horizon Natural Resource Damage Assessment Trustees, 2016). Like with other large marine oil spills, this 2010 spill prompted considerable research to characterize the biological effects of acute crude oil exposure in aquatic animals (Incardona, 2017). In fish, crude oil toxicity has been attributed to the induction of impaired cardiovascular development in embryos (Incardona et al., 2006, 2004; Jung et al., 2013; Khursigara et al., 2017; Perrichon et al., 2021; Sørhus et al., 2016; Xu et al., 2017). Other acute studies have also shown that oil exposure can produce teratogenic effects such as abnormal craniofacial, spinal, and muscular development in both freshwater and marine teleosts (de Soysa et al., 2012b; Incardona et al., 2014; Incardona and Scholz, 2016; Philibert et al., 2019). Despite the importance of these studies, little is known about the long-term consequences of abnormal embryonic or larval development to the morphology and fitness of animals late in life.
Although oil is most acutely toxic in fish during the embryonic and larval life stages, the tissues of adult fish exposed to oil show altered gene expression patterns suggestive of impaired reproduction (Pilcher et al., 2014; Whitehead et al., 2012). These studies suggest the potential for persistent impacts of transient oil exposure, including effects that may span generations. Given our growing understanding of epigenetic modes of inheritance in fish (Labbé et al., 2017), there has been increasing interest in the transgenerational effects of crude oil exposure. Recent studies on Macondo oil from the Deepwater Horizon oil spill have begun to characterize the multigenerational impacts of parental crude oil exposure (Bautista and Burggren, 2019; Jasperse et al., 2019a, 2019b; Vignet et al., 2015). Traditionally, most studies have investigated the effects of both parental lineages on transgenerational inheritance or have preferentially focused solely on the maternal input. Collectively, these studies have shown somatic, developmental, behavioral, and reproductive effects when both parents are exposed to oil (Bautista and Burggren, 2019; Jasperse et al., 2019a, 2019b). However, there is growing interest in the role of paternal modes of epigenetic inheritance (Bautista et al., 2020). In zebrafish (Danio rerio), it was found that the methylome is responsible for epigenetic gene regulation, and that inheritance occurs via the sperm and not the oocyte (Jiang et al., 2013). Overall, these studies demonstrate the persistence of altered pheno- and epigenotypes across multiple generations, although additional work is needed to assess the maternal versus paternal modes of transgenerational inheritance. Also, assessments of biological effects in progeny should be extended to include adult phenotypes and other fitness level endpoints.
Traditional assessments of commonly observed adult phenotypes such as altered spinal and craniofacial development have been conducted in a presence-absence manner in oil toxicity studies (de Soysa et al., 2012a; Hansen et al., 2019; Raimondo et al., 2014). However, these studies do not account for the variance in total body shape for a treatment group. An alternate approach that would allow for the identification of fine-scale changes in body shape across treatments would be through the usage of landmark-based geometric morphometrics (GM). In past studies, landmark-based GM has been used to identify patterns of altered morphology in response to marine pollution in wild populations of fish (Gharred et al., 2020), bivalves (Reyna et al., 2019; Scalici et al., 2017), and gastropods (Primost et al., 2016). Only a few laboratory studies have utilized GM techniques to assess the effects of aquatic toxicants in fish; although morphology has been shown altered by PCB or TCDD exposure in fathead minnows (Coulter et al., 2019) and zebrafish (Chollett et al., 2014), respectively. Although two-dimensional (2D) GM application to environmental toxicity is less common in the context of transgenerational studies, this technique could provide a powerful approach to identify subtle changes in animal structure due to toxicant exposure.
In the current study, we investigated the effects of parental crude oil exposure on adult offspring for two generations using an assessment of swimming performance and GM techniques to quantify variation in body shape changes. The ubiquitous nGoM marsh resident, the Gulf killifish (Fundulus grandis), was used for this study as it is an ecologically relevant model. The Gulf killifish was one of the many nGOM marsh species that experienced oiling during its peak spawning period following the Deepwater Horizon oil spill. Male and female Gulf killifish were exposed for 36–44 days to sublethal concentrations of Macondo-252 light sweet crude oil. These fish were then used to generate one of four lineages of F1 fish using a full-matrix mating design with gametes from control and oil-exposed F0 adults. The F1 generation was not exposed to oil directly at any time following fertilization. This F1 generation was then used to create an F2 generation of fish that also was never exposed directly to oil. We noticed upon maturation that morphological abnormalities were present. For this reason, we assessed the aerobic swimming capacity for both F1 and F2 fish. We additionally conducted ad hoc landmark-based GM and subsequent multivariate generalized linear models (GLM) to determine the effects of parental crude oil exposure on (1) fish morphology and (2) patterns of maternal or paternal transmission of altered phenotypes in two generations.
Section snippets
Generation of water accommodated fraction for crude oil exposures
A custom-built recirculation system was constructed, similar to that described in Carls et al. (2000) and Kennedy and Farrell (2005), to produce water accommodated fraction (WAF) of crude oil by mechanical agitation. The recirculating system consisted of a 1000-L tank with a polytetrafluoroethylene (PTFE) liner, a water pump at the tank base, and a PTFE pipe connected to the pump. The upweller pipe had a compartment where oiled sintered glass beads (Siporax®, 15 mm, Sera) in a PTFE-mesh bag
PAC analysis
Fish were exposed to WAF of Macondo crude oil with a mean TPAC50 concentration (mean ± SEM) of 68.1 ± 15.9 µg L − 1 (n = 6 composite water samples; range: 20.44 µg L−1–123.21 µg L−1 [PAC]). TPAC50 analysis identified that 92.54% ± 0.49% were composed of alkylated PAC homologs (Supplemental Data Table S3). On average, naphthalenes, fluorenes, and phenanthrenens/anthracenes contributed 65.0 ± 0.6% to the TPAC50 concentration. Adult fish were exposed to treatment water composed of 2–3 ring PACs
Crude oil exposure causes transgenerational changes in swimming capacity and body shape
The current study aimed to identify the effects of parental oil exposure on swimming capacity and morphology across multiple generations of progeny. We subsequently found evidence that F0 paternal oil exposure resulted in fish with altered body shape and decreased aerobic swimming capabilities. These results also showed the persistence of altered phenotypes and impaired aerobic swimming performance across two generations following parental (F0) oil exposure, particularly when paternally exposed.
CRediT authorship contribution statement
Chelsea Hess: Project administration, Conceptualization, Methodology, Formal analysis, Investigation, Visualization, Writing – original draft. Lauren Little: Investigation. Charles Brown: Investigation. Michael Kaller: Formal analysis, Writing – review & editing. Fernando Galvez: Funding acquisition, Project administration, Methodology, Conceptualization, Resources, Writing – review & editing, Supervision.
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have influenced the work reported in this paper.
Acknowledgements
This research was funded by grants from the National Science Foundation (EF-0723771) and the National Academies of Sciences Gulf Research Program to FG and a training grant by the Louisiana Sea Grant Undergraduate Research Opportunities Program to LL and FG. Training in geometric morphometric analysis via Transmitting Science was funded by Louisiana Sea Grant and Louisiana State University BioGrads research awards to CH. iPad provided by Becky Carmichael, Ph.D., with Louisiana State University
References (61)
Life-history patterns of the salt-marsh killifish Fundulus heteroclitus (L.) introduced in the estuary of the guadalquivir river (South West Spain)
Estuar. Coast. Shelf Sci.
(1989)Paternal exposures: impact on reproductive and developmental outcome. An overview
Pharmacol. Biochem. Behav.
(1996)- et al.
Developmental effects in fish embryos exposed to oil dispersions – The impact of crude oil micro-droplets
Mar. Environ. Res.
(2019) - et al.
Defects in cardiac function precede morphological abnormalities in fish embryos exposed to polycyclic aromatic hydrocarbons
Toxicol. Appl. Pharmacol.
(2004) - et al.
Developmental toxicity of 4-ring polycyclic aromatic hydrocarbons in zebrafish is differentially dependent on AH receptor isoforms and hepatic cytochrome P4501A metabolism
Toxicol. Appl. Pharmacol.
(2006) - et al.
The influence of heart developmental anatomy on cardiotoxicity-based adverse outcome pathways in fish
Aquat. Toxicol.
(2016) - et al.
Sperm, but not oocyte, DNA methylome is inherited by zebrafish early embryos
Cell
(2013) - et al.
Sustained impairment of respiratory function and swim performance following acute oil exposure in a coastal marine fish
Aquat. Toxicol.
(2017) - et al.
Geologically distinct crude oils cause a common cardiotoxicity syndrome in developing zebrafish
Chemosphere
(2013) - et al.
Ion homeostasis and interrenal stress responses in juvenile Pacific herring, Clupea pallasi, exposed to the water-soluble fraction of crude oil
J. Exp. Mar. Biol. Ecol.
(2005)
Cardiac function and survival are affected by crude oil in larval red drum, Sciaenops ocellatus
Sci. Total Environ.
Exposure to deepwater horizon weathered crude oil increases routine metabolic demand in chub mackerel, Scomber japonicus
Mar. Pollut. Bull.
Transgenerational inheritance of neurobehavioral and physiological deficits from developmental exposure to benzo[a]pyrene in zebrafish
Toxicol. Appl. Pharmacol.
Epigenetics in fish gametes and early embryo
Aquaculture
Cardio-respiratory function during exercise in the cobia, Rachycentron canadum: the impact of crude oil exposure
Comp. Biochem. Physiol. Part C Toxicol. Pharmacol.
Effects of deepwater horizon crude oil exposure, temperature and developmental stage on oxygen consumption of embryonic and larval mahi-mahi (Coryphaena hippurus)
Aquat. Toxicol.
Field-collected crude oil, weathered oil and dispersants differentially affect the early life stages of freshwater and saltwater fishes
Sci. Total Environ.
Developmental toxicity of Louisiana crude oil-spiked sediment to zebrafish
Ecotoxicol. Environ. Saf.
A multilevel response approach reveals the Asian clam Corbicula largillierti as a mirror of aquatic pollution
Sci. Total Environ.
Hydrocarbons (jet fuel JP-8) induce epigenetic transgenerational inheritance of obesity, reproductive disease and sperm epimutations
Reprod. Toxicol.
The interactive effects of feeding and exercise on oxygen consumption, swimming performance and protein usage in juvenile rainbow trout (Oncorhynchus mykiss)
J. Exp. Biol.
Parental stressor exposure simultaneously conveys both adaptive and maladaptive larval phenotypes through epigenetic inheritance in the zebrafish (Danio rerio)
J. Exp. Biol.
Parental transgenerational epigenetic inheritance related to dietary crude oil exposure in Danio rerio
J. Exp. Biol.
Be careful with your principal components
Evolution (N Y)
Swimming performance of sockeye salmon (Oncorhynchus nerka) in relation to fatigue time and temperature
J. Fish. Res. Board Can.
The respiratory metabolism and swimming performance of young sockeye salmon
J. Fish. Res. Board Can.
Exposure of pacific herring to weathered crude oil: assessing effects on ova
Environ. Toxicol. Chem.
Embryonic exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin impairs prey capture by zebrafish larvae
Environ. Toxicol. Chem.
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