Electronic supplementary materialSperfeld Wacker – Hydrobiologia

Appendix B:Additional information for Offspring diet treatments experiment.

Fig. B1Juvenile mass-specific growth rates (d-1) of D. magna offspring raised on pure Synechococcuselongatus(SYN). Mothers have been raised on Scenedesmusobliquus (SCE), Nannochloropsislimnetica (NAN), and a food mixture containing 95% SYN, 2.5% SCE, and 2.5% NAN supplemented with cholesterol (MF+Ch). Mean ± SD are presented for n = 3 (except for NAN, n = 2). There are no significant differences among maternal diet treatments (one-way ANOVA: F2,5 = 0.89, p = 0.47). Replicates started with 14-15 juveniles in 500 mL food suspension and were conducted only until day 4 as animals already showed first signs of mortality on this poor food; dry mass was determined at the beginning and day 4 for calculation of mass-specific growth rates.

Table B1 Fatty acid concentrations (µg mg C-1) of phytoplankton (Food) and Daphnia magna offspring (Neos). Phytoplankton used as food in experiments: Synechococcuselongatus (SYN), Scenedesmusobliquus (SCE), Nannochloropsislimnetica (NAN), and a food mixture containing 95% SYN, 2.5% SCE, and 2.5% NAN supplemented with cholesterol (MF+Ch).

SYN / MF+Ch / SCE / NAN
Food / Food / Neos / Food / Neos / Food / Neos
 SFA / 42.0
(2.6) / 41.3
(2.5) / 39.2
(4.1) / 24.8 (2.3) / 35.6
(4.6) / 38.0
(3.1) / 48.1
(6.9)
 MUFA / 50.9
(1.9) / 49.7
(1.8) / 76.2
(3.0) / 17.3
(2.3) / 26.5
(3.8) / 42.5
(2.7) / 58.6
(7.7)
ALA (18:33) / n.d. / 1.6
(0.3) / 7.1
(1.5) / 48.3
(3.4) / 72.4
(5.2) / 2.5
(0.4) / 5.6
(0.1)
SDA (18:43) / n.d. / 0.16
(0.02) / 1.3
(0.4) / 5.0
(0.3) / 6.1
(0.5) / n.d. / 0.87
(0.15)
EPA (20:53) / n.d. / 1.4
(0.4) / 5.0
(0.5) / n.d. / n.d. / 70.4
(5.8) / 92.3
(12.5)
LIN
(18:26) / n.d. / 0.43
(0.06) / 2.9
(0.8) / 10.7
(0.4) / 15.4
(1.0) / 5.0
(0.7) / 8.0
(0.6)
ARA (20:46) / n.d. / 0.18
(0.05) / 1.7
(0.3) / n.d. / n.d. / 9.0
(0.9) / 11.6
(1.4)
3 PUFA / n.d. / 3.1
(0.5) / 13.3
(2.4) / 53.3
(3.7) / 78.5
(5.7) / 73.0
(6.1) / 98.8
(12.4)
6 PUFA / n.d. / 0.72
(0.11) / 5.7
(2.2) / 12.7
(0.4) / 19.5
(1.6) / 16.1
(1.5) / 23.5
(2.0)
TFA / 98.8
(4.8) / 101.6
(4.9) / 138.9
(12.6) / 141.3
(10.2) / 197.1
(18.2) / 175.4
(8.8) / 237.0
(29.7)
3/6 ratio / - / 4.3
(0.2) / 2.6
(0.6) / 4.2
(0.3) / 4.0
(0.04) / 4.5
(0.3) / 4.2
(0.2)

Footnotes Table B1: Data of Food are means (± SD) of five subsamples taken at different days of the maternal and offspring diet treatments. Data of Neos are means of duplicate measurements (± min./max.);n.d., not detected. SFA represent the sum of all saturated fatty acids ≥ 14 carbon atoms; MUFA represent the sum of all monounsaturated fatty acids ≥ 16 carbon atoms; PUFA represent the sum of all polyunsaturated fatty acids with at least two double bonds and ≥ 18 carbon atoms: α-linolenic acid (ALA), stearidonic acid (SDA), eicosapentaenoic acid (EPA), linoleic acid (LIN), arachidonic acid (ARA); TFA (total fatty acids) represent the sum of all fatty acids.

Fatty acid concentrations of the Food MF+Ch were calculated from monoculture measurements at the same sampling date. Offspring were freshly hatched neonates from the third clutch of D. magna females pre-cultured upon SCE, NAN, and MF+Ch.The measured fatty acid amounts of each sample were normalized to independently determined POC amounts of phytoplankton and to the carbon content of D. magna’s offspring using a previously determined carbon to dry mass conversion factor of 0.43 µg carbon per µg dry mass (n=147). Preliminary measurements have shown no difference in carbon content of D. magna grown on diets that differed substantially in PUFA composition and variation around the mean of the conversion factor was rather small (coefficient of variation ±3.6%).