Do distributions of diamondoid hydrocarbons accumulated in oil-contaminated fish tissues help to identify the sources of oil?

Identifying the sources of environmental oil contamination can be challenging, especially for oil in motile organisms such as fish. Lipophilic hydrocarbons from oil can bioaccumulate in fish adipose tissue and potentially provide a forensic "fingerprint" of the original oil. Herein, diamondoid hydrocarbon distributions were employed to provide such fingerprints. Indices produced from diamondoids were used to compare extracts from fish adipose tissues and the crude and fuel oils to which the fish were exposed under laboratory conditions. A suite of 20 diamondoids was found to have bioaccumulated in the dietary-exposed fish. Cross-plots of indices between fish and exposure oils were close to the ideal 1:1 relationship. Comparisons with diamondoid distributions of non-exposure oils produced overall, but not exclusively, weaker correlations. Linear Discriminatory Analysis on a combined set of 15 diamondoid and bicyclane molecular ratios was able to identify the exposure oils, so a use of both compound classes is preferable.

Dataset on aquatic ecotoxicity predictions of 2697 chemicals, using three quantitative structure-activity relationship platforms.

Empirical and in silico data on the aquatic ecotoxicology of 2697 organic chemicals were collected in order to compile a dataset for assessing the predictive power of current Quantitative Structure Activity Relationship (QSAR) models and software platforms. This document presents the dataset and the data pipeline for its creation. Empirical data were collected from the US EPA ECOTOX Knowledgebase (ECOTOX) and the EFSA (European Food Safety Authority) report "Completion of data entry of pesticide ecotoxicology Tier 1 study endpoints in a XML schema - database". Only data for OECD recommended algae, daphnia and fish species were retained. QSAR toxicity predictions were calculated for each chemical and each of six endpoints using ECOSAR, VEGA and the Toxicity Estimation Software Tool (T.E.S.T.) platforms. Finally, the dataset was amended with SMILES, InChIKey, pKa and logP collected from webchem and PubChem.

Fish Fingerprinting: Identifying Crude Oil Pollutants using Bicyclic Sesquiterpanes (Bicyclanes) in the Tissues of Exposed Fish.

In the present study, we investigated the possibility of identifying the source oils of exposed fish using ratios of bicyclic sesquiterpane (bicyclane) chemical biomarkers. In the event of an oil spill, identification of source oil(s) for assessment, or for litigation purposes, typically uses diagnostic ratios of chemical biomarkers to produce characteristic oil "fingerprints." Although this has been applied in identifying oil residues in sediments, water, and sessile filtering organisms, so far as we are aware this has never been successfully demonstrated for oil-exposed fish. In a 35-day laboratory trial, juvenile Lates calcarifer (barramundi or Asian seabass) were exposed, via the diet (1% w/w), to either a heavy fuel oil or to Montara, an Australian medium crude oil. Two-dimensional gas chromatography with high-resolution mass spectrometry and gas chromatography-mass spectrometry were then used to measure selected ratios of the bicyclanes to examine whether the ratios were statistically reproducibly conserved in the fish tissues. Six diagnostic bicyclane ratios showed high correlation (r2 > 0.98) with those of each of the two source oils. A linear discriminatory analysis model showed that nine different petroleum products could be reproducibly discriminated using these bicyclane ratios. The model was then used to correctly identify the bicyclane profiles of each of the two exposure oils in the adipose tissue extracts of each of the 18 fish fed oil-enriched diets. From our initial study, bicyclane biomarkers appear to show good potential for providing reliable forensic fingerprints of the sources of oil contamination of exposed fish. Further research is needed to investigate the minimum exposure times required for bicyclane bioaccumulation to achieve detectable concentrations in fish adipose tissues and to determine bicyclane depuration rates once exposure to oil has ceased. Environ Toxicol Chem 2023;42:7-18. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Gut Microbiome as a Potential Biomarker in Fish: Dietary Exposure to Petroleum Hydrocarbons and Metals, Metabolic Functions and Cytokine Expression in Juvenile Lates calcarifer.

The gut microbiome of fish contains core taxa whose relative abundances are modulated in response to diet, environmental factors, and exposure to toxicogenic chemicals, influencing the health of the host fish. Recent advances in genomics and metabolomics have suggested the potential of microbiome analysis as a biomarker for exposure to toxicogenic compounds. In this 35-day laboratory study, 16S RNA sequencing and multivariate analysis were used to explore changes in the gut microbiome of juvenile Lates calcarifer exposed to dietary sub-lethal doses of three metals: vanadium (20 mg/kg), nickel (480 mg/kg), and iron (470 mg/kg), and to two oils: bunker C heavy fuel oil (HFO) (1% w/w) and Montara, a typical Australian medium crude oil (ACO) (1% w/w). Diversity of the gut microbiome was significantly reduced compared to negative controls in fish exposed to metals, but not petroleum hydrocarbons. The core taxa in the microbiome of negative control fish comprised phyla Proteobacteria (62%), Firmicutes (7%), Planctomycetes (3%), Actinobacteria (2%), Bacteroidetes (1%), and others (25%). Differences in the relative abundances of bacterial phyla of metal-exposed fish were pronounced, with the microbiome of Ni-, V-, and Fe-exposed fish dominated by Proteobacteria (81%), Firmicutes (68%), and Bacteroidetes (48%), respectively. The genus Photobacterium was enriched proportionally to the concentration of polycyclic aromatic hydrocarbons (PAHs) in oil-exposed fish. The probiotic lactic acid bacterium Lactobacillus was significantly reduced in the microbiota of fish exposed to metals. Transcription of cytokines IL-1, IL-10, and TNF-a was significantly upregulated in fish exposed to metals but unchanged in oil-exposed fish compared to negative controls. However, IL-7 was significantly downregulated in fish exposed to V, Ni, Fe, and HFOs. Fish gut microbiome exhibits distinctive changes in response to specific toxicants and shows potential for use as biomarkers of exposure to V, Ni, Fe, and to PAHs present in crude oil.

Multivariate analysis of otolith microchemistry can discriminate the source of oil contamination in exposed fish.

The uptake of metals into the aragonite lattice of the fish otolith (ear-bone) has been used for decades as a historical record of exposure to metals in polluted environments. The relative abundance of two metals in particular, Ni and V, are used in forensic chemical analysis of crude oils to assist in confirming its origin. In this study we investigate the potential for metal accumulation in otoliths to act as a biomarker of exposure to crude oil. Using a 33-day static-renewal laboratory trial design, 56 juvenile Lates calcarifer (commonly known as Asian seabass or barramundi) were fed diets enriched with V (20 mg/kg), Ni (500 mg/kg), Fe (500 mg/kg), and two crude oils with distinctly different metals profiles: a heavy fuel oil (1% w/w) and a typical Australian medium crude (1% w/w). Fish exposed to crude oils showed Ba and Al retained in otoliths in a dose-dependent manner, but fish fed V-, Ni- and Fe-enriched diets showed no metal increase in otoliths, indicating that V, Ni and Fe are not incorporated into the otolith of L. calcarifer via dietary exposure. For crude oils, incorporation into otolith for many metals is likely limited due to porphyrin casing reducing their bioavailability. Principal components analysis (PCA) and subsequent linear discriminatory analysis (LDA) of selected otolith metals demonstrated that, even despite large variability in the metal abundances detected in otolith between individuals within the test groups (cv = 1.00), it is possible to discriminate between fish exposed to different crude oils using multivariate analysis of their otolith microchemistry.

Discriminating source of oil contamination in teleost fish, Lates calcarifer, using multivariate analysis of a suite of physiological and behavioral biomarkers.

The release of petroleum hydrocarbons into the environment from natural seeps, well blowouts, pipeline leaks, shipping accidents and deliberate tank washing poses an ongoing threat to marine ecosystems. Distinguishing the source of oil contamination in exposed biota can be relatively straightforward if samples of the oil are available but, in their absence, such discrimination in fish poses a major challenge. The use of physiological and behavioral biomarker analysis provides a useful tool to describe sub-lethal effects of toxicant exposure. In this study we describe the responses of 12 biomarkers in Lates calcarifer (Asian seabass) following a 33-day dietary exposure (1%w/w) to heavy fuel oil (HFO) and to Montara, a typical Australian medium crude oil (MCO). Principal components analysis was used to differentiate between fish exposed to HFO from those exposed to MCO. Inferences can be made about the composition of an oil from the biomarker profiles produced in exposed fish.

PLASMA BIOCHEMISTRY PROFILES OF WILD WESTERN TIGER SNAKES (NOTECHIS SCUTATUS OCCIDENTALIS) BEFORE AND AFTER SIX MONTHS OF CAPTIVITY.

Urban wildlife often suffer poorer health than their counterparts living in more pristine environments due to exposure to anthropogenic stressors such as habitat degradation and environmental contamination. As a result, the health of urban versus nonurban snakes might be assessed by differences in their plasma biochemistries. We compared the plasma profiles of western tiger snakes (Notechis scutatus occidentalis) from a heavily urbanized wetland and a natural, nonurbanized wetland. Despite the urbanized snakes having lower body mass index, we found no significant difference between the plasma profiles of the two populations. We collected snakes from each population and kept them in captivity for 6 mo, providing them with stable conditions, uncontaminated (exempt from heavy metals and pesticides) food and water, and lowered parasite intensity in an attempt to promote better health through depuration. After captivity, snakes experienced a significant improvement in body mass index and significant changes in their plasma profiles. Snakes from the natural wetland initially had more variation of DNA damage; mean concentration of DNA damage in all snakes slightly decreased, but not significantly, after captivity. We present the plasma biochemistry profiles from western tiger snakes both before and after captivity and suggest a period of removal from natural stressors via captivity may offer a more reliable result of how plasma profiles of healthy animals might appear.

Assessing the effects of textile leachates in fish using multiple testing methods: From gene expression to behavior.

The textile industry, while of major importance in the world economy, is a toxic industry utilizing and emitting thousands of chemical substances into the aquatic environment. The aim of this project was to study the potentially harmful effects associated with the leaching of chemical residues from three different types of textiles: sportswear, children's bath towels, and denim using different fish models (cell lines, fish larvae and juvenile fish). A combination of in vitro and in vivo test systems was used. Numerous biomarkers, ranging from gene expression, cytotoxicity and biochemical analysis to behavior, were measured to detect effects of leached chemicals. Principle findings indicate that leachates from all three types of textiles induced cytotoxicity on fish cell lines (RTgill-W1). Leachates from sportswear and towels induced mortality in zebrafish embryos, and chemical residues from sportswear reduced locomotion responses in developing larval fish. Sportswear leachate increased Cyp1a mRNA expression and EROD activity in liver of exposed brown trout. Leachates from towels induced EROD activity and VTG in rainbow trout, and these effects were mitigated by the temperature of the extraction process. All indicators of toxicity tested showed that exposure to textile leachate can cause adverse reactions in fish. These findings suggested that chemical leaching from textiles from domestic households could pose an ecotoxicological threat to the health of the aquatic environment.

Risk Assessment of Pesticide Mixtures in Australian Rivers Discharging to the Great Barrier Reef.

Rivers discharging to the Great Barrier Reef carry complex pesticide mixtures. Here we present a first comprehensive ecotoxicological risk assessment using species sensitivity distributions (SSDs), explore how risk changes with time and land use, and identify the drivers of mixture risks. The analyzed data set comprises 50 different pesticides and pesticide metabolites that were analyzed in 3741 samples from 18 river and creek catchments between 2011 and 2016. Pesticide mixtures were present in 82% of the samples, with a maximum of 23 pesticides and a median of five compounds per sample. Chemical-analytical techniques were insufficiently sensitive for at least seven pesticides (metsulfuron-methyl, terbutryn, imidacloprid, clothianidin, ametryn, prometryn, and thiamethoxam). The classical mixture concepts of concentration addition and independent action were applied to the pesticide SSDs, focusing on environmental threshold values protective for 95% of the species. Both concepts produced almost identical risk estimates. Mixture risk was therefore finally assessed using concentration addition, as the sum of the individual risk quotients. The sum of risk quotients ranges between 0.05 and 122 with a median of 0.66. An ecotoxicological risk (i.e., a sum of individual risk quotients exceeding 1) was indicated in 38.5% of the samples. Sixteen compounds accounted for 99% of the risk, with diuron, imidacloprid, atrazine, metolachlor, and hexazinone being the most important risk drivers. Analysis of land-use patterns in catchment areas showed an association between sugar cane farming and elevated risk levels, driven by the presence of diuron.