To investigate changes in the proportion of plant macrofossils vs. coarse grained inorganic sediments entering

the lake, dried bulk sediment samples were sieved at 600 μm. BMS-387032 in vivo The samples were then submerged in water and the floating (organic macrofossil) and sinking (inorganic, coarse grains) fractions separated. The organic macrofossil fraction was dried, weighed and expressed as a percentage of the original total sample mass. The ratio between total carbon and total nitrogen (TC:TN) may be used as an indicator of whether the organic matter is primarily aquatic (TC:TN < 10) or terrestrial (TC:TN > 10) in origin (Meyers and Teranes, 2001). Hence, TC:TN ratios can be used to study changes in the source of the organic material present in the sediment. TC and TN were measured at 0.5 cm intervals using 20–60 mg of sediment with a Macro Vario elemental analyser. The TC and TN contents of the organic macrofossils were also measured. Total sulphur (TS) was measured at 5 cm intervals E7080 solubility dmso using approximately 2 g dried sediment with a LECO CNS 2000 analyser. Diatoms are one of the most commonly used biological indicators of aquatic ecosystem changes (Smol, 2008). They are highly sensitive and respond rapidly to changes in

their environment (e.g. light, nutrients, pH, salinity, sediment supply and temperature; Smol and Stoermer, 2010). Diatoms were analysed at 0.5 cm intervals using standard methods (Battarbee et al., 2001). At least 400 valves were counted per sample, using phase contrast and oil immersion at 1000× magnification on a Dolichyl-phosphate-mannose-protein mannosyltransferase Zeiss Z20 light microscope. The relative abundance

of all species (including unidentified forms) was recorded as a percentage of the total number of valves counted (Battarbee et al., 2001). Taxonomy was principally based on sub-Antarctic (Van de Vijver et al., 2002), Antarctic (Roberts and McMinn, 1999) and Australian taxonomic literature (Vyverman et al., 1995 and Hodgson et al., 1997). All taxa were photographed and are archived, including taxonomic data, with K. Saunders. Species occurring with ≥1% relative abundance were included in this study. Separate constrained hierarchical cluster analyses (CONISS; Grim, 1987) were undertaken on the sedimentological (water content, plant macrofossil, TC, TN, TS) and diatom data to determine the timing of the most significant splits in the data, in particular whether the most significant split coincided with the introduction of rabbits. The broken stick model was used to determine the number of significant splits (Bennett, 1996). This identifies a zone boundary as significant if the explained variance of the zonation exceeds the variance of a zonation in a random dataset with the same parameters (i.e. n and total variance the same as in the actual dataset; Bennett, 1996). These analyses were performed in R version 15.