F₁ group

The number of male and female fish used in each F1 treatment arm. In total there were 16 fish (4.6 fish/L) in each tank. Mortality prior to the intervention resulted in the loss of one fish from Tank 2 in the 5–5 arm and mortality during the intervention resulted in the loss of five fish from the 5–5 and 5–60 arms (parental 5 mg diet).</p

Reproductive consequences of dietary intake.

<p>Fertility changes are shown for group 3; for a comparison of the three groups see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0166394#pone.0166394.s002" target="_blank">S2 Fig</a>. The (A) breeding success, (B) clutch size, and (C) fertilization rate is plotted for group 3. Using incrosses within the 5 mg treatment arm (F: 5 mg x M: 5 mg) as the reference for comparison, the effect of the 60 mg treatment was observed using incrosses (F: 60 mg x M: 60 mg) and outcrosses of males (F: 5 mg x M: 60 mg) and females (F: 60 mg x M: 5 mg). The values given represent the odds ratio (A-C), rate ratio (D-F), or incidence rate ratio (G-I) from an average of three pairs from each of three tanks in four spawning experiments. Standard errors were calculated with respect to the tank clusters and are presented as a ± 95% CI. Statistically significant differences are noted as * p ≤ 0.05, ** p ≤ 0.01, and **** p ≤ 0.0001.</p

F₀ groups.

<p>The number of male and female fish used in each F₀ treatment arm. All the fish were of the same strain, with the groups born a month apart from one another. We aimed to have 20 fertile fish in three tanks within each treatment arm in order to be able to detect a breeding odds ratio of 0.4 with 75% power [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0166394#pone.0166394.ref025" target="_blank">25</a>]. On average there were 20 fish (5.7 fish/L) in each tank for group 2 and 3. In group 1, fertility screening only allowed 12 fish (3.4 fish/L) to be allocated into each tank. Tank 3 in group 1 has 10 fish due to mortality prior to the intervention.</p

Phenotypic changes resulting from nutrient availability.

<p>Gross phenotypic changes are shown only for group 3, for a comparison of the three groups see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0166394#pone.0166394.s001" target="_blank">S1 Fig</a>. (A) Food consumption over the eight weeks of the dietary intervention for the 5 mg arm (meal 1) and the 60 mg arm (meals 1, 2, and 3). The number of <i>Artemia</i> remaining following feeding is given as a percentage of the number dispensed to each tank, note the scale begins at 96%. (B) The BMI derived from the weights and lengths of every fish in the 5 and 60 mg treatment arms before and after the dietary intervention. The statistical differences are noted between the treatment arms (solid lines) and within each treatment arm (dotted lines), before and after the diet. (C) The total distance travelled in 30 seconds of swimming for the populations of fish in the 5 and 60 mg treatment arms at the start (week 1) and the end (week 8) of the diet. Values represent the mean ± SEM from the three tanks within each group. Statistically significant differences are noted as **** p ≤ 0.0001.</p

RNA sequencing revealed diet-induced differences in egg transcript deposition.

<p>(A) The differentially expressed genes are shown in terms of the number of genes significantly altered in each direction. The significance cut-off was–log(<i>p</i> = 0.005) = 2.3. The top 15 biological processes overrepresented within the list of genes significantly (B) downregulated and (C) upregulated in the 60 mg arm are given as bars with the size indicating the significance of the enrichment.</p

Parental diet affects F₁ survival.

<p>(A) The survival of pooled offspring from incrosses within the 5 and 60 mg treatment arms over their maturation. (B) The growth of the offspring as determined by the length of 5 randomly selected fish from each tank from each parental treatment arm (n = 60 per condition) over the course of their development. (C) The sex ratio of the offspring. Values represent the mean ± SEM from three tanks within each group. Statistically significant differences are noted as * p ≤ 0.05 and **** p ≤ 0.0001.</p

Parental diet affects F₁ physical activity.

<p>(A) Food consumption over the eight weeks of the dietary intervention for the F₁ 5 mg arms (meal 1) and the F₁ 60 mg arms (meals 1, 2, and 3). The number of <i>Artemia</i> remaining following feeding is given as a percentage of the number dispensed to each tank, note the scale begins at 75%. (B) The BMI derived from the weights and lengths of every fish in the 5 and 60 mg treatment arms before and after the dietary intervention. (C) The total distance travelled in 30 seconds of swimming for the populations of fish in the 5 and 60 mg treatment arms at the start (week 1) and the end (week 8) of the diet. Values represent the mean ± SEM from the three tanks within each group. Statistically significant differences are noted as * p ≤ 0.05 and **** p ≤ 0.0001.</p

Summary of demographic features of survey participants.

Summary of demographic features of survey participants.</p

Summary of participant perceptions of the impact of strained ICU capacity.

Summary of participant perceptions of the impact of strained ICU capacity.</p

Summary of the proportion of time health care workers perceived their ICU is strained due to capacity issues.

Summary of the proportion of time health care workers perceived their ICU is strained due to capacity issues.</p