Primers used to amplify 16S rDNA and integron components.

<p>Primers used to amplify 16S rDNA and integron components.</p

Antibiotic resistance integrons in feces from captive rock wallabies.

<p>(<b>A</b>) A wild brush-tailed rock-wallaby meets an animal released from a captive breeding program (on right, with radio tracking collar). Photo Credit: Hugh McGregor (<b>B</b>) Schematic maps of integron cassette arrays recovered from 14 of 29 captive wallabies. Numbers of wallabies with each array combination are shown. Red diamonds – the primary integron recombination site, <i>attI1</i>; red circles – gene cassette recombination sites, <i>attC</i>; broad arrows – genes showing direction of transcription. Gene symbols are as follows: <i>aadA</i> genes encode aminoglycoside adenyltransferases that confer resistance to streptomycin & spectinomycin; <i>qac</i> confers resistance to quaternary ammonium compounds, <i>dfr</i> genes encode dihydrofolate reductases that confer resistance to trimethoprim, <i>gcuF</i> unknown function (<i>5</i>).</p

Class 1 integrons identified in captive rock wallabies from Waterfall Springs.

<p>Presence of the class 1 integron-integrase gene, <i>intI1</i>, as determined by PCR with primers HS464/HS463a (A) and the identity of integron gene cassettes as determined by sequence analysis of cassette arrays amplified with primers HS458/HS459 (B). Two distinct cassette arrays were detected in some wallaby samples (C).</p

Modelled (FSR_mod) and observed (FSR_obs) fertilization success for each urchin pair under acidified conditions (pH 7.6 and 7.8), and parameters from Control observations (pH 8.1) used in modelling FSR_mod at lowered pH levels.

<p>FSR_{50 Control} = 50% of maximum fertilization success in Controls; <i>F</i>_{50 Control} = sperm concentration that generates 50% of maximum fertilization success in Controls. Sperm data from each male in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053118#pone-0053118-t002" target="_blank">Table 2</a> were used in modelling FSR_mod. No females were spawned for male A.</p

Seawater parameters for the three different pH treatments.

<p>pH<b>_NBS</b>, temperature (T), salinity (Sal) and total alkalinity (A_T) were measured directly and used to compute partial pressure levels of carbon dioxide (<i>p</i>CO₂) and seawater saturation states for calcite and aragonite (Ω_Ca and Ω_Ar respectively) using CO2SYS. Means ± S.E.</p

Scatterplots for observed (FSR_obs) versus modelled (FSR_mod) fertilization success for pH 7.8 (A) and 7.6 (B).

<p>Regression analyses revealed a significant relationship between observed (independent) and modelled fertilization (dependent) for pH 7.8 (<i>P</i> = 0.012, r² = 0.336), but not for pH 7.6 (<i>P</i> = 0.413, r² = 0.042).</p

Ocean acidification effects on sperm speed and percent sperm motility for each male <i>Heliocidaris erythogramma</i>.

<p>Significant differences in parameters between sites (<i>P</i>≤0.05) are shown in bold.</p

Impacts of ocean acidification on sperm motility and sperm swimming speed in <i>Heliocidaris erythrogramma.</i>

<p>Proportion of mean (±S.E.) motile sperm (A) and sperm speed (B) at different levels of ocean acidification (pH mediated by CO₂ addition). Lower case letters indicate significantly different groups at <i>p</i> = 0.05 (Tukey’s test). (C) Mean logarithmic response ratios (±95% CI) of effects of ocean acidification on percent motility and sperm speed (n = 19).</p

Two-way ANOVA for percent motility (A) and sperm speed (B) of <i>Heliocidaris erythrogramma</i> across different pH treatments (fixed) and males (random).

<p>Significant effects (<i>P</i>≤0.05) are shown in bold.</p

Two-way ANOVA for observed (FSR_obs; A) and modelled fertilization success (FSR_mod; B) of <i>Heliocidaris erythrogramma</i> across different pH treatments (fixed) and males (random).

<p>Significant effects (P≤0.05) are shown in bold.</p