Transcriptomic and Histological Analysis of the Greentail Prawn (Metapenaeus bennettae) Following Light Crude Oil Exposure.

Oil spills pose a significant threat to marine biodiversity. Crude oil can partition into sediments where it may be persistent, placing benthic species such as decapods at particular risk of exposure. Transcriptomic and histological tools are often used to investigate the effects of hydrocarbon exposure on marine organisms following oil spill events, allowing for the identification of metabolic pathways impacted by oil exposure. However, there is limited information available for decapod crustaceans, many of which carry significant economic value. In the present study, we assess the sublethal impacts of crude oil exposure in the commercially important Australian greentail prawn (Metapenaeus bennettae) using transcriptomic and histological analyses. Prawns exposed to light, unweathered crude oil "spiked" sediments for 90 h were transferred to clean sediments for a further 72 h to assess recovery. Chemical analyses indicated that polycyclic aromatic hydrocarbons increased by approximately 65% and 91% in prawn muscle following 24 and 90 h of exposure, respectively, and significantly decreased during 24- and 72-h recovery periods. Transcriptomic responses followed an exposure and recovery pattern with innate immunity and nutrient metabolism transcripts significantly lowered in abundance after 24 h of exposure and were higher in abundance after 72 h of recovery. In addition, transcription/translation, cellular responses, and DNA repair pathways were significantly impacted after 24 h of exposure and recovered after 72 h of recovery. However, histological alterations such as tubule atrophy indicated an increase in severity after 24 and 72 h of recovery. The present study provides new insights into the sublethal impacts of crude oil exposure in greentail prawns and identifies molecular pathways altered by exposure. We expect these findings to inform future management associated with oil extraction activity and spills. Environ Toxicol Chem 2022;41:2162-2180. © 2022 John Wiley & Sons Ltd. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Monitoring sublethal changes in fish physiology following exposure to a light, unweathered crude oil.

Biomarkers are frequently used to determine the exposure of fish to petroleum hydrocarbons following an oil spill. These biomarkers must be chosen carefully if they are to be used to determine sublethal toxic impacts as well as oil exposure. Many commonly used biomarkers relate to the metabolism of high molecular weight, typically pyrogenic, polycyclic aromatic hydrocarbons (PAHs), which are not abundant in unweathered crude oil. The goal of this study was to compare the efficacy of different biomarkers, including histological examination and transcriptomic profiling, in showing exposure to oil and the potential for sublethal toxic impacts. To achieve these goals, subadults/adults of the spotted dragonet (Repomucenus calcaratus) were exposed to a representative light, unweathered Australian oil for 96 h, so that the physiological changes that occur with exposure could be documented. Fish were then transferred to clean sediment for 90 h to quantify recovery. Biomarker changes, including PAH metabolites, 7-ethoxyresorufin O-deethylase (EROD), and histopathology, are presented in this work. In addition, a de novo transcriptome for the spotted dragonet was assembled, and differential transcript abundance was determined for the gill and liver of petroleum-exposed fish relative to a control. Increased levels of some biliary phenanthrene metabolites were seen throughout the exposure period. EROD levels showed modest, but not significant, increases. Transcriptomic differences were noted in the abundances of transcripts with a role in inflammation, primary metabolism and cardiac function. The patterns of transcript abundance in the gill and the liver changed in a manner that reflected exposure and recovery. The histology showed elevated prevalence of lesions, most notably vacuolization in liver and heart tissue, multi-organ necrosis, and lamellar epithelial lifting and telangiectasia in the gill. These findings suggest that short-term exposures to low molecular weight PAHs could elicit changes in the health of fish that are well predicted by the transcriptome. Furthermore, when light oil is released into the environment, exposure and subsequent risk would be better estimated using phenanthrene metabolite levels rather than EROD. This study also adds to the weight of evidence that exposure to low molecular weight PAHs may cause cardiac problems in fish. Further study is needed to determine the impact of these changes on reproductive capacity, long-term survival, and other population specific parameters.

Transcriptomic, lipid, and histological profiles suggest changes in health in fish from a pesticide hot spot.

Barramundi (Lates calcarifer) were collected at the beginning (1st sampling) and end (2nd sampling) of the wet season from Sandy Creek, an agriculturally impacted catchment in the Mackay Whitsundays region of the Great Barrier Reef catchment area, and from Repulse Creek, located approximately 100 km north in Conway National Park, to assess the impacts of pesticide exposure. Gill and liver histology, lipid class composition in muscle, and the hepatic transcriptome were examined. The first sample of Repulse Creek fish showed little tissue damage and low transcript levels of xenobiotic metabolism enzymes. Sandy Creek fish showed altered transcriptomic patterns, including those that regulate lipid metabolism, xenobiotic metabolism, and immune response; gross histological alterations including lipidosis; and differences in some lipid classes. The second sampling of Repulse Creek fish showed similar alterations in hepatic transcriptome and tissue structure as fish from Sandy Creek. These changes may indicate a decrease in health of pesticide exposed fish.

The environmental impact of sewage and wastewater outfalls in Antarctica: An example from Davis station, East Antarctica.

We present a comprehensive scientific assessment of the environmental impacts of an Antarctic wastewater ocean outfall, at Davis station in East Antarctica. We assessed the effectiveness of current wastewater treatment and disposal requirements under the Protocol on Environmental Protection to the Antarctic Treaty. Macerated wastewater has been discharged from an outfall at Davis since the failure of the secondary treatment plant in 2005. Water, sediment and wildlife were tested for presence of human enteric bacteria and antibiotic resistance mechanisms. Epibiotic and sediment macrofaunal communities were tested for differences between sites near the outfall and controls. Local fish were examined for evidence of histopathological abnormalities. Sediments, fish and gastropods were tested for uptake of sewage as measured by stable isotopes of N and C. Escherichia coli carrying antibiotic resistance determinants were found in water, sediments and wildlife (the filter feeding bivalve Laternula eliptica). Fish (Trematomus bernacchii) within close proximity to the outfall had significantly more severe and greater occurrences of histopathological abnormalities than at controls, consistent with exposure to sewage. There was significant enrichment of 15N in T. bernacchii and the predatory gastropod Neobuccinum eatoni around the outfall, providing evidence of uptake of sewage. There were significant differences between epibiotic and sediment macrofaunal communities at control and outfall sites (<1.5 km), when sites were separated into groups of similar habitat types. Benthic community composition was also strongly related to habitat and environmental drivers such as sea ice. The combined evidence indicated that the discharge of wastewater from the Davis outfall is causing environmental impacts. These findings suggest that conditions in Antarctic coastal locations, such as Davis, are unlikely to be conducive to initial dilution and rapid dispersal of wastewater as required under the Protocol on Environmental Protection to the Antarctic Treaty. Current minimum requirements for wastewater treatment and disposal in Antarctica are insufficient to ameliorate these risks and are likely to lead to accumulation of contaminants and introduction of non-native microbes and associated genetic elements. This new understanding suggests that modernised approaches to the treatment and disposal of wastewater are required in Antarctica. The most effective solution is advanced levels of wastewater treatment, which are now possible, feasible and a high priority for installation. As a direct outcome of the study, a new advanced treatment system is being installed at Davis, effectively avoiding environmental risks.

Application of a quantitative histological health index for Antarctic rock cod (Trematomus bernacchii) from Davis Station, East Antarctica.

A quantitative Histological Health Index (HHI) was applied to Antarctic rock cod (Trematomus bernacchii) using gill, liver, spleen, kidney and gonad to assess the impact of wastewater effluent from Davis Station, East Antarctica. A total of 120 fish were collected from 6 sites in the Prydz Bay region of East Antarctica at varying distances from the wastewater outfall. The HHI revealed a greater severity of alteration in fish at the wastewater outfall, which decreased stepwise with distance. Gill and liver displayed the greatest severity of alteration in fish occurring in close proximity to the wastewater outfall, showing severe and pronounced alteration respectively. Findings of the HHI add to a growing weight of evidence indicating that the current level of wastewater treatment at Davis Station is insufficient to prevent impact to the surrounding environment. The HHI for T. bernacchii developed in this study is recommended as a useful risk assessment tool for assessing in situ, sub-lethal impacts from station-derived contamination in coastal regions throughout Antarctica.

Tracking spatial distribution of human-derived wastewater from Davis Station, East Antarctica, using δ15N and δ13C stable isotopes.

Stable isotope ratios, δ15N and δ13C were effectively used to determine the geographical dispersion of human derived sewage from Davis Station, East Antarctica, using Antarctic rock cod (Trematomus bernacchii). Fish within 0-4 km downstream of the outfall exhibited higher δ15N and δ13C values relative to reference sites. Nitrogen in particular showed a stepped decrease in δ15N with increasing distance from the discharge point by 1-2‰. Stable isotopes were better able to detect the extent of wastewater contamination than other techniques including faecal coliform and sterol measures. Uptake and assimilation of δ15N and δ13C up to 4 km from the outfall adds to growing evidence indicating the current level of wastewater treatment at Davis Station is not sufficient to avoid impact to the surrounding environment. Isotopic assimilation in T. bernacchii is a viable biomarker for investigation of initial sewage exposure and longer term monitoring in the future.

Direct evidence of histopathological impacts of wastewater discharge on resident Antarctic fish (Trematomus bernacchii) at Davis Station, East Antarctica.

During the 2009/2010 summer, a comprehensive environmental impact assessment (EIA) of the wastewater discharge at Davis Station, East Antarctica was completed. As part of this, histological alteration of gill and liver tissue in Antarctic Rock-cod (Trematomus bernacchii) from four sites along a spatial gradient from the wastewater outfall were assessed. All fish within 800 m of the outfall exhibited significant histological changes in both tissues. Common pathologies observed in fish closest to the outfall include proliferation of epithelial cells with associated secondary lamellar fusion in the gills and multifocal granulomata with inflammation and necrosis as well as cysts in the liver. Fish from sites >800 m from the outfall also exhibited alterations but to a lesser degree, with prevalence and severity decreasing with increasing distance from the outfall. This study highlights the value of histopathological investigations as part of EIAs and provides the first evidence of sub-lethal alteration associated with wastewater discharge in East Antarctica.

Optometry-based general population survey of pupil ruff atrophy and ocular hypertension.

BACKGROUND: To evaluate and describe the pupil ruff changes and relationship to intraocular pressure, pseudoexfoliation syndrome and glaucoma status in an optometric population in New Zealand. DESIGN: Prospective cross-sectional survey of an optometric population. PARTICIPANTS: Six hundred and twenty subjects over 50 years old routinely attending the participating optometry practices. Exclusion criteria included previous intraocular surgery, ophthalmic laser, uveitis, angle closure and secondary glaucoma. METHODS: Multicentre study involving 11 optometry practices in the Wellington region, New Zealand. The pupillary ruff and associated gonioscopy findings of study participants were graded based on the previously published Pupil Ruff Atrophy grading system. Parameters evaluated include pupillary ruff absence and abnormality, pseudoexfoliation material and trabecular meshwork pigmentation. MAIN OUTCOME MEASURES: Correlations between intereye Pupil Ruff Atrophy grading differences and inter-eye intraocular pressure and cup:disc ratio differences. RESULTS: Six hundred and twenty subjects were included, with a mean age of 62.2 ± 9.1 years and mean intraocular pressure of 14.8 ± 3.4 mmHg. Four hundred and fourteen (66.8%) had bilateral pupil ruff changes and 12 (1.5%) had pseudoexfoliation. Inter-eye intraocular pressure asymmetry was significantly correlated with amount of missing pupillary ruff (r = 0.111; P = 0.022) and trabecular meshwork pigmentation (r = 0.147; P = 0.002). Inter-eye cup:disc ratio asymmetry was not correlated with any of the Pupil Ruff Atrophy grading parameters. CONCLUSIONS: Asymmetry of pupillary ruff absence and trabecular meshwork pigmentation was correlated with intraocular pressure asymmetry (but not with cup:disc ratio asymmetry) in a general optometric population setting in New Zealand.

Emergency contraception: knowledge and perceptions in a university population.

PURPOSE: The purpose of this study was to examine knowledge, attitudes, and behaviors regarding emergency contraception (EC) in university men and women aged 18-21. DATA SOURCES: Data sources included responses to a 25-item questionnaire and an 8-item demographic survey completed anonymously at a public site on campus. Ninety-seven university students participated in the study. Participants were asked to respond to questions relating to knowledge, attitudes, and behaviors regarding EC, perceived worthiness, objections, sources of information about EC, preferred birth-control method and usage, and perceptions of their personal risk of unintended pregnancy. CONCLUSIONS: Many respondents considered unintended pregnancy to be a major problem and considered EC a worthy option in the event of method failure or unprotected intercourse. While most participants were aware that there was a postcoital method of contraception, confusion existed between EC and RU-486 (the abortion pill). Almost half (49.5%) believed that EC was the same as RU-486. There was an association between advanced prescription for EC and its likelihood of use. Most women would be significantly more likely to use EC if they had a prescription on hand. Of the women who were less likely to choose EC, 100% indicated they would feel embarrassed or judged when asking for it. Only 34% of those women who have had a gynecological exam in the past 12 months had discussed EC with their provider. IMPLICATIONS FOR PRACTICE: Advanced practice nurses need to incorporate EC into preventive health counseling for both men and women. Providing women with an advanced prescription increases the likelihood that women will use EC.