The effects of petroleum hydrocarbon contamination on selected intertidal macrophytes and meiofauna.
The effects of bunker fuel oil on the growth of A. marina, B. gymnorrhiza and R. mucronata were investigated in glasshouse and field experiments. The effects of oil on community structure in micro-organisms were also investigated in microcosm glasshouse experiments. The differences in oil tolerance of the three mangroves were compared in propagule and sediment oiled treatments and growth monitored for 13 months under glasshouse conditions. In propagule oiled treatments, various portions of the propagule were coated with oil. In the sediment oiled treatments, 50ml oil were added to the sediment in each pot. In oiled treatments, plant height, number of leaves and chlorophyll content were significantly reduced in all species compared to the control. In A. marina and R. mucronata, oiling resulted in growth malformations such as abnormal phyllotaxy and deformity of leaves and stems. The effects of oil on root growth were investigated in rhizotrons for 245 and 409 days respectively. In oiled treatments, root growth rate, length and volume were significantly reduced in all species. In A. marina and B. gymnorrhiza oil increased root diameter. In another series of experiments, PAH accumulation in roots and leaves of the three species were determined in one year old seedlings subjected to oiling for 21 days. The concentrations of 15 PAHs in roots and leaves were determined by gas chromatography / mass spectrometry. The highest total concentration of PAHs was accumulated in oiled roots of A. marina (44,045.9μg/kg), followed by B. gymnorrhiza (10,280.4μg/kg) and R. mucronata (6,979.1μg/kg). In oiled treatments, the most common PAHs in roots of all species were fluorene and acenaphthene (two rings), phenanthrene and anthracene (three rings), pyrene and chrysene (four rings) and benzo[a]pyrene (five rings). In the leaves of all species in oiled treatments, the common PAHs were naphthalene and acenapthene (two rings) and phenanthrene (three rings). To test for living and dead root tip cells and to compare the effects of oil on cell ultrastructure in roots and leaves of the three species, one year old seedlings were subjected to a control and sediment oiled treatments for seven days. Control root tips, stained with fluorescein diacetate, exhibited green fluorescence in living cells of the meristematic and conducting tissue in all species. Oiled root tips, stained with propidium iodide, exhibited red fluorescence, indicating cell death or dead cells. Transmission electron micrographs revealed that oil damaged cell ultrastructure in root tips and leaves in all species. Anatomical changes induced by oil included, disorganization of cells in the root cap, epidermis and meristem. Oil also induced loss of cell contents and destruction of organelles in root tissue. Oil damaged chloroplasts and cell organelles in spongy mesophyll and palisade cells of leaves. To compare the effects of oil on the ability of the three species to tolerate salinity, healthy one year old seedlings were subjected to 10% and 50% seawater in control and sediment oiled treatments for 12 months. In the oiled treatments, 200ml oil were added to the soil in each pot. Oil significantly reduced growth in the 50% seawater treatment in all species. Results suggested that oil reduces salt tolerance in the three species. The effects of oil on salt secretion in A. marina were investigated by subjecting one year old seedlings to sediment oiling treatments at 0%, 10% and 50% seawater for three weeks. Sodium accumulated in the leaves of oiled seedlings at 10% and 50% seawater. The effects of oil on salt secretion in A. marina in the light and dark were compared in one year old seedlings subjected to oiling treatments for seven days. Sodium accumulated in the leaves of oiled seedlings in the light and dark within 11 hours. Oil reduced secretion rates of Na⁺, K⁺, Ca²⁺ and Mg²⁺ in all treatments. The effects of oil on species abundance, richness and community structure of soil micro-organisms were determined by subjecting microcosms to oiling treatments with or without fertiliser for four weeks. In the oiled treatments, 15ml oil and 5ml/L fertiliser were added to 200g soil. Fertiliser consisted of 4% N, 2% P and 5% K. Nematodes were extracted after the experimental period and identified to genus or species level. Oil significantly reduced species abundance and richness. Oil also eliminated sensitive species and altered the abundance of dominant species thereby altering the free living nematode community structure. Addition of fertiliser increased richness and dominant species in oiled treatments. The effects of oil coating on leaves and internodes on growth of the three mangroves were investigated in field experiments for 48 weeks. Oiling of the leaves resulted in leaf abscission and decreased leaf production in all species. The effects of sediment oiling (at a dose of 5Lmˉ²) on the three species were also investigated in a field study for 53 weeks. In A. marina, oil caused adventitious roots to develop on the stem, about 10-15 cm above the soil surface after 38 weeks of treatment. In oiled treatments, plant mortality occurred after 53 weeks in all three species. The ability of B. gymnorrhiza and R. mucronata to exclude PAHs from sensitive root tissues probably accounted for the higher oil tolerance than A. marina. The capacity of the species to adapt to residual oil contamination by increasing root diameter (A. marina and B. gymnorrhiza), producing adventitious roots (A. marina), increasing root/shoot ratio (R. mucronata) and abscising oiled leaves (all species) probably contributed to oil tolerance.
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