Immunofluorescence detection and localization of B[a]P and TCDD in earthworm tissues

An immunohistochemical method using antibodies against polycyclic aromatic hydrocarbons (PAHs) and dioxins was developed on frozen tissue sections of the earthworm Eisenia andrei exposed to environmentally relevant concentrations of benzo[a]pyrene (B[a]P) (0.1, 10, 50 ppm) and 2,3,7,8-tetrachlorodibenzo-para-dioxin (TCDD) (0.01, 0.1, 2 ppb) in spiked standard soils. The concentrations of B[a]P and TCDD in E. andrei exposed to the same conditions were also measured using analytical chemical procedures. The results demonstrated that tissues of worms exposed to even minimal amount of B[a]P and TCDD reacted positively and specifically to anti-PAHs and -dioxins antibody. Immunofluorescence revealed a much more intense staining for the gut compared to the body wall; moreover, positively immunoreactive amoeboid coelomocytes were also observed, i.e. cells in which we have previously demonstrated the occurrence of genotoxic damage. The double immunolabelling with antibodies against B[a]P/TCDD and the lysosomal enzyme cathepsin D demonstrated the lysosomal accumulation of the organic xenobiotic compounds, in particular in the cells of the chloragogenous tissue as well as in coelomocytes, involved into detoxification and protection of animals against toxic chemicals. The method described is timesaving, not expensive and easily applicable.

Benthic megafauna and CO2 bubble dynamics observed by underwater photography during a controlled sub-seabed release of CO2

In 2012, a controlled sub-seabed release of carbon dioxide (CO2) was conducted in Ardmucknish Bay, a shallow (12 m) coastal bay on the west coast of Scotland. During the experiment, CO2 gas was released 12 m below the seabed for 37 days, causing significant disruption to sediment and water carbonate chemistry as the gas passed up through the sediment and into the overlying water. One of the aims of the study was to investigate how the impacts caused by leakage from geological CO2 Capture and Storage (CCS) could be detected and quantified in the context of natural heterogeneity and dynamics. To do this underwater photography was used to analyze (i) the benthic megafaunal response to the CO2 release and (ii) the dynamics of the CO2 bubble streams, emerging from the seabed into the overlying water column. The frequently observed megafauna species in the study area were Virgularia mirabilis (Cnidaria), Turritella communis (Mollusca), Asterias rubens (Echinodermata), Pagurus bernhardus (Crustacea), Liocarcinus depurator (Crustacea), and Gadus morhua (Osteichthyes). No discernable abnormal behavior was observed for these megafauna, in any of the zones investigated, during or after the CO2 release. Time-lapse photography revealed that the intensity and presence of the CO2 bubble plume was affected by the tides, with the most active bubbling seen at low tides and the larger hydrostatic pressure at high tide suppressing CO2 bubbling from the seabed.