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Coral growth anomalies (GAs) are tumor-like protrusions that are detrimental to coral health, affecting both the coral skeleton and soft tissues. These lesions are increasingly found throughout the tropics and are commonly associated with high human population density, yet little is known about the molecular pathology of the disease. Here, we investigate the metabolic impacts of GAs through 1H nuclear magnetic resonance (NMR) metabolomics in Porites compressa tissues from a site of high disease prevalence (Coconut Island, Hawaii). We putatively identified 18 metabolites (8.1% of total annotated features) through complementary 1H and 1H–13C heteronuclear single quantum correlation NMR data that increase confidence in pathway analyses and may bolster future coral metabolite annotation efforts. Extract yield was elevated in both GA and unaffected (normal tissue from a diseased colony) compared to reference (normal tissue from GA-free colony) samples, potentially indicating elevated metabolic activity in GA-impacted colonies. Relatively high variation in metabolomic profiles among coral samples of the same treatment (i.e., inter-colony variation) confounded data interpretation, however, analyses of paired GA and unaffected samples identified 73 features that differed between these respective metabolome types. These features were largely annotated as unknowns, but 1-methylnicotinamide and trigonelline were found to be elevated in GA samples, while betaine, glycine, and histamine were lower in GA samples. Pathway analyses indicate decreased choline oxidation in GA samples, making this a pathway of interest for future targeted studies. Collectively, our results provide unique insights into GA pathophysiology by showing these lesions alter both the absolute and relative metabolism of affected colonies and by identifying features (metabolites and unknowns) and metabolic pathways of interest in GA pathophysiology going forward.

Mercury has been identified as one of the most serious environmental threats to the well-being of wildlife in the southeastern United States (Facemire et al. 1995). The prevalence of Hg in aquatic species has also prompted concerns for the health of subsistence fishermen and the general population who regularly consume fish. This concern is evident in the fact that 76% of fish consumption advisories in the United States in 2003 were due at least in part to Hg, for a total of 5,289,020 hectares of lakes and 1,234,127 river miles [U.S. Environmental Protection Agency (EPA) 2004]. The toxic effects of Hg have been demonstrated in mammals, birds, and fish and include neurotoxicity, impaired growth and development, reduced reproductive success, liver and kidney damage, and immunomodulation (Wiener et al. 2003; Zelikoff et al. 1994). Reptiles are prominent members of ecosystems and often have life history characteristics that make them vulnerable to Hg accumulation (e.g., long life span, high trophic level, aquatic habitat). Despite the fact that many of these reptilian species also have a tenuous conservation status, there are currently few data on the toxicity of Hg in this taxon. One such example is the sea turtle, of which all species are classified as either threatened or endangered. Many anthropogenic factors have been implicated in the decline in sea turtle populations, including directed harvest for food and trade, fisheries bycatch, and degradation of nesting beach habitat (Lutcavage et al. 1997). However, the role of chemical pollutants in marine turtle health is largely unknown. It is therefore important to understand the risk that contaminants pose to the general health and immunologic function of sea turtles because these effects could also impact the survival of their populations. Several studies have measured Hg levels in tissues from juvenile and adult sea turtles (Anan et al. 2001; Davenport and Wrench 1990; Day et al. 2005; Godley et al. 1999; Gordon et al. 1998; Maffucci et al. 2005; Orvik 1997; Presti 1999; Sakai et al. 1995, 2000a, 2000b; Storelli et al. 1998, 2005; Wang 2005). However, relatively few studies have assessed health parameters in sea turtles in relation to environmental contaminants (Keller et al. 2004, 2005, 2006a, 2006b; Lutcavage et al. 1995; Peden-Adams et al. 2002, 2003; Podreka et al. 1998), and these studies have focused primarily on organic contaminants rather than metals. Balazs and Pooley (1991) suggested that environmental contaminants are a possible factor contributing to the development of the viral disease fibropapillomatosis in sea turtles by reducing immune function. Some locations in Florida (Indian River Lagoon and Florida Bay) exhibit up to 70% prevalence of this disease, and these environments also have elevated levels of Hg (Ache et al. 2000; Cantillo et al. 1999; Trocine and Trefry 1996). In the present study we investigated the relationship of blood Hg levels in loggerhead sea turtles (Caretta caretta) off the coasts of South Carolina, Georgia, and northern Florida to various immunologic and general health parameters. Using an integrated approach, correlative field data collection from wild-caught loggerhead sea turtles were combined with laboratory in vitro exposure of loggerhead lymphocytes to methylmercury (MeHg).

The contents from thick-billed murre (Uria lomvia) eggs collected at four Alaskan colonies in 2002 were analyzed for organic contaminants and carbon δ¹³C) and nitrogen (δ¹⁵N) stable isotopes. Contaminant concentrations in the eggs varied from below detection limits to 230 ng g⁻¹ wet mass for 4,4'-DDE in one egg from St Lazaria Island in the Gulf of Alaska. Eggs from this colony generally contained higher levels of contaminants and exhibited significantly different patterns compared to eggs from the Bering and Chukchi seas. Stable isotope values also varied geographically; however, these differences appeared to be related to differences in C and N baselines in the food webs instead of differences in prey. Contaminant and stable isotope correlations were inconclusive, suggesting that better information on regional food web differences and differential offloading of contaminants and stable isotopes to the eggs must be obtained before these kinds of data can be fully incorporated into seabird egg contaminant monitoring programs.

A highly sensitive and selective method has been developed for the determination of methylmercury in biological specimens and NIST Standard Reference Materials (SRMs). The procedure involves microwave extraction with acetic acid, followed by derivatization and headspace solid-phase microextraction (SPME) with a polydimethylsiloxane (PDMS)-coated silica fiber. Optimization of conditions including gas chromatograph injection temperature, microwave extraction power and microwave extraction time are presented. The identification and quantification (via the method of standard additions) of the extracted compounds is carried out by capillary gas chromatography with inductively coupled plasma mass spectrometric detection (GC-ICP-MS) using a unique heated interface that was designed for this work. The SPME-GC-ICP-MS method was validated for the determination of methylmercury (MeHg) concentrations in a variety of biological Standard Reference Materials (SRMs), ranging from 13.2 ng g−1 in SRM 1566b Oyster Tissue, to 397 ng g−1 in SRM 1946 Lake Superior Fish Tissue. Additionally, this method was applied to the determination of MeHg in seabird eggs (common murres, Uria aalge and thick-billed murres, Uria lomvia) collected from colonies on Little Diomede and Bogoslof islands in the Bering Sea and Saint Lazaria Island in the Gulf of Alaska and cryogenically banked in the Marine Environmental Specimen Bank. The results obtained demonstrate that SPME-GC-ICP-MS is a sensitive technique for the determination of methylmercury at trace and ultra-trace levels in a variety of natural matrices with high reproducibility and accuracy. In all instances, the sample-to-sample variability was typically 2% relative standard deviation (RSD) and the method detection limit for methylmercury was 4.2 pg g−1 (as Hg), based on a 0.5 g tissue sample of SRM 1566b Oyster Tissue.