Multi-domain probiotic consortium as an alternative to chemical remediation of oil spills at coral reefs and adjacent sites

Background Beginning in the last century, coral reefs have suffered the consequences of anthropogenic activities, including oil contamination. Chemical remediation methods, such as dispersants, can cause substantial harm to corals and reduce their resilience to stressors. To evaluate the impacts of oil contamination and find potential alternative solutions to chemical dispersants, we conducted a mesocosm experiment with the fire coral Millepora alcicornis, which is sensitive to environmental changes. We exposed M. alcicornis to a realistic oil-spill scenario in which we applied an innovative multi-domain bioremediator consortium (bacteria, filamentous fungi, and yeast) and a chemical dispersant (Corexit® 9500, one of the most widely used dispersants), to assess the effects on host health and host-associated microbial communities. Results The selected multi-domain microbial consortium helped to mitigate the impacts of the oil, substantially degrading the polycyclic aromatic and n-alkane fractions and maintaining the physiological integrity of the corals. Exposure to Corexit 9500 negatively impacted the host physiology and altered the coral-associated microbial community. After exposure, the abundances of certain bacterial genera such as Rugeria and Roseovarius increased, as previously reported in stressed or diseased corals. We also identified several bioindicators of Corexit 9500 in the microbiome. The impact of Corexit 9500 on the coral health and microbial community was far greater than oil alone, killing corals after only 4 days of exposure in the flow-through system. In the treatments with Corexit 9500, the action of the bioremediator consortium could not be observed directly because of the extreme toxicity of the dispersant to M. alcicornis and its associated microbiome. Conclusions Our results emphasize the importance of investigating the host-associated microbiome in order to detect and mitigate the effects of oil contamination on corals and the potential role of microbial mitigation and bioindicators as conservation tools. Chemical dispersants were far more damaging to corals and their associated microbiome than oil, and should not be used close to coral reefs. This study can aid in decision-making to minimize the negative effects of oil and dispersants on coral reefs.

Effects of sub-lethal concentrations of corexit 9500® dispersant on growth and condition factor of juvenile Clarias gariepinus (Burchell 1822), Rivers state, Nigeria

The effects of sub-lethal concentrations of Corexit 9500 on growth and condition factor of juveniles of Clarias gariepinus was investigated after 21days, in 2018. 160 fishes were used as test organisms with a loading of 10/treatment and were exposed to 0.00, 0.0125, 0.025 and 0.05 ml/L concentrations of Corexit 9500. Treatments were in triplicate and Complete Randomised Design was adopted. The growth in length and weight of C. gariepinus was significantly higher in the control than the test treatments (P<0.05). The increase in weight of the control was significantly different from the three treatments. Temperature was not significantly different (P>0.05); however, the DO of control was significantly higher (P < 0.05) than that of the treatments, with the lowest DO in the 0.05ml/l treatment. pH was acidic for all treatments including the control but was more acidic with increasing concentration of Corexit 9500. The pH value for each treatment was significantly different (P<0.05). The conductivity was significantly different from each other (P<0.05) with higher values at the 0.025 ml/L concentrations. The condition factor (K) decreased with increasing toxicant concentration and was significantly different (P<0.05) at 0.0125ml/L and at Weeks 2 and 3 of 0.05 ml/L. Corexit 9500 dispersants should be appropriately assessed before deployment, and their use near fish breeding grounds should be avoided. Key words: Clarias gariepinus, dispersant, growth, physicochemical parameters, sub-lethal

The Interactive Effects of Crude Oil and Corexit 9500 on Their Biodegradation in Arctic Seawater

ABSTRACT The risk of petroleum spills coupled with the potential application of chemical dispersants as a spill response strategy necessitates further understanding of the fate of oil and dispersants and their interactive effects during biodegradation. Using Arctic seawater mesocosms amended with either crude oil, Corexit 9500, or both together, we quantified the chemical losses of crude oil and Corexit 9500 and identified microbial taxa implicated in their biodegradation based on shifts in the microbial community structure over a 30-day time course. Chemical analyses included total petroleum hydrocarbons (TPH), n-alkanes, branched alkanes, and polycyclic aromatic hydrocarbons (PAHs) for oil loss and the surfactant components dioctyl sodium sulfosuccinate (DOSS), Span 80, Tween 80, Tween 85, and the DOSS metabolite ethylhexyl sulfosuccinate (EHSS) for Corexit loss. Changes to the microbial communities and identification of key taxa were determined by 16S rRNA gene amplicon sequencing. The nonionic surfactants of Corexit 9500 (Span 80 and Tweens 80 and 85) biodegraded rapidly, dropping to below the limits of detection within 5 days and prior to any detectable initiation of oil biodegradation. This resulted in no observable suppression of petroleum biodegradation in the presence of Corexit compared to that of oil alone. In contrast, biodegradation of DOSS was delayed in the presence of oil, based on the prolonged presence of DOSS and accumulation of the degradation intermediate EHSS that did not occur in the absence of oil. Microbial analyses revealed that oil and Corexit enriched different overall microbial communities, with the presence of both resulting in a community composition that shifted from one more similar to that of Corexit only to one reflecting the oil-only community over time, in parallel with the degradation of predominantly Corexit and then oil components. Some microbial taxa (Oleispira, Pseudofulvibacter, and Roseobacter) responded to either oil or Corexit, suggesting that some organisms may be capable of utilizing both substrates. Together, these findings reveal interactive effects of crude oil and Corexit 9500 on chemical losses and microbial communities as they biodegrade, providing further insight into their fate when copresent in the environment. IMPORTANCE Chemical dispersants such as Corexit 9500 are commonly used in oil spill response and are currently under consideration for use in the Arctic, where their fate and effects have not been well studied. This research was performed to determine the interactive effects of the copresence of crude oil and Corexit 9500 on the degradation of components from each mixture and the associated microbial community structure over time in Arctic seawater. These findings will help yield a better understanding of the biodegradability of dispersant components applied to an oil spill, the temporal microbial community response to dispersed oil, and the fundamental microbial ecology of organic contaminant biodegradation processes in the Arctic marine environment.

The Impact of the Deepwater Horizon Oil Spill upon Lung Health—Mouse Model-Based RNA-Seq Analyses

We used a transcriptomic approach to interrogate the effects of a saline-accommodated fraction from the Macondo 252 well (MC252) oil and Corexit dispersants on lung tissue. Wild-type C57BL/6 male and female mice were exposed on days 0, 7 and 13 by oropharyngeal aspiration to saline accommodated fractions (SAF) of crude oil from the Macondo (MC252) well, Corexit 9500, Corexit 9527, 9500+oil and 9527+oil or a saline solution as the vehicle control. These treatments did not cause overt toxicity, with the exception of the Corexit exposures which caused brief weight loss after the first exposure. On day 14, total RNA was isolated from the left lung for RNA-seq analyses. KEGG-pathway-based differential expression revealed that Corexit 9527 elicited the strongest changes involving the upregulation of 19 KEGG pathways (FDR < 0.10), followed by Corexit 9500 with the upregulation of seven pathways (FDR < 0.10). As an important signature, pathways related to a response to DNA damage (e.g., p53 signaling and mismatch repair) dominate those upregulated by Corexit 9527 and Corexit 9500. In addition, pro-inflammatory pathways (e.g., cytokine-cytokine receptor interaction, IL-17 signaling pathway and TNF signaling pathways) were upregulated selectively in oil-treated male mice. Surprisingly, oil + dispersant combinations caused lesser effects than the individual treatments at the transcriptomic level. Overall, these findings support potential genotoxicity, inflammation and cell death due to dispersant or oil exposures. Similar exposures to lung tumor bearing K-RasLA1 mice provided evidence for tumor promotion by oil and Corexit dispersant treatments. Our mouse RNA-seq analyses may be relevant to the pulmonary health hazards of MC252 oil and dispersants experienced in exposed populations.

Histo-toxicity of Corexit 9500 Dispersant on Gill and Liver of Clarias gariepinus (Burchell 1822)

The aim of this study was to assess the chronic toxicity of Corexit 9500 on histology of liver and gill of juveniles of Clarias gariepinus. Range finding tests were conducted over a 96-hr period after acclimatization of the test organisms in the C.P. Powell laboratory in the University of Port Harcourt, Nigeria. The test organisms 10/treatment were exposed to the following concentrations of Corexit 9500; 0.00 ml/L, 0.0125 ml/L, 0.025 ml/L and 0.05 ml/L in triplicate. Liver and gill were excised from fish every week for histological assay using standard methods. The LC50 at 96 hrs was 0.115 ml/L. The growth in length and weight of C. gariepinus was significantly higher in the control than the test treatments (p<0.05). Hyperplasias, necrosis of epithelial cells, inflammation of the secondary gill lamella and hypertrophic primary gill lamella were observed in the exposed gills. Hepatic lesions in the liver tissues of the fishes exposed to Corexit 9500 were characterized by hyperplasia, narrowing of the central vein and vacoulations. The alterations observed in the liver and gill tissues indicated that Corexit 9500 may have interfered with transamination and metabolic processes, with the possibility of affecting the physiological functions of the fish in an aquatic environment.