Depression-like behavior of HAB animals in the tail suspension test (TST).

<p>Animals fed with blue-green algae diet hat a strongly reduced immobility, indicating depression-like behavior, compared to standard fed animals. This effect was even more pronounced in animals fed with the ¹⁵N isotope (*p<0.05; **p<0.01).</p

Food consumption of algae and bacteria diets of HAB/NAB/LAB dams and pups before weaning.

<p>Consumption of blue-green algae diet of HAB dams and their pups and consumption of bacteria diet of HAB/NAB/LAB dams and their pups before weaning (post natal days (PND) 1–24); ¹⁴N diet 14N; ¹⁵N-enriched diet.</p

Depression-like behavior in the tail suspension test (TST).

<p>High (HAB), normal (NAB) and low (LAB) anxious animals were fed with ¹⁴N and/or ¹⁵N enriched bacteria diets. <b>A</b>) No behavioral changes due to diet <i>per se</i> were found for animals fed with bacteria diet compared to the animals of the respective lines of the HAB/NAB/LAB standard breeding of the same generation (dotted lines and indicated in <b>B</b>) as percentage difference of the standard breeding). However, ¹⁵N fed HAB animals showed a strongly reduced immobility, indicating depression-like behavior, compared to ¹⁴N fed HABs (**p<0.01). B) This differed significantly from the standard HAB/NAB/LAB breeding (*p<0.05). Due to division by zero, no value is given for ¹⁴N fed LAB animals in relation to the standard breeding: values are ¹⁴N LAB 0.53±0.3 sec vs. standard LAB 4.06±1.5 sec.</p

Incorporation rate of ¹⁵N in (A) plasma and (B) brain proteins.

<p>In 5, 14 and 28 day old animals (PND 5, 14, 28) bacteria diet feeding provides a significantly higher incorporation of ¹⁵N in plasma proteins compared to blue-green algae diet; only at the age of 56 days, blue-green algae fed diet animals show the same ¹⁵N incorporation. In the cerebellum, a faster incorporation in adolescence at PND 14 reached significance (**p<0.01; *p<0.05).</p

Anxiety-related behavior and locomotion of HAB animals on the elevated plus maze (EPM).

<p>No behavioral changes due to the blue-green algae diet fed to the animals were found.</p

Analysis of expression differences by MS analysis and RT-PCR.

<p><b>A</b>) ESI mass spectrum of the ¹⁴N LAB (red) and ¹⁵N HAB (blue) isotope forms of the Hint2 peptide ISQAEEDDQQLLGHLLLVAK in cerebellar tissue. The isotopologue patterns of the ¹⁴N and ¹⁵N peptide signals (m/z) were used for relative quantification with ProRata. <b>B</b>) Extracted ion chromatograms for the Hint2 peptide, extracted from A, showing an upregulation of Hint2 in HAB mice. <b>C</b>) Analysis of Hint2 transcripts in ¹⁵N-labeled HAB and ¹⁴N LAB cerebellar cDNA by RT-PCR, normalized by HRSP12 expression. Hint2 expression in HABs was 2.5-fold higher compared to LABs (*p<0.05). No smoothing was applied.</p

Body weight after weaning of animals fed with bacteria, blue-green algae and standard diet.

<p>Compared to standard fed HAB animals (orange dotted line), blue-green algae diet fed animals were significantly heavier (green line) and bacteria fed animals lighter (red line) (**p<0.01 blue-green algae vs. control ++p<0.01 bacteria vs. control).</p

Analysis of expression differences by MS analysis.

<p><b>A</b>) ESI mass spectrum of the ¹⁴N LAB (red) and ¹⁵N HAB (blue) isotope forms of the GLO1 peptide GFGHIGIAVPDVYSACK in cerebellar tissue. The isotopologue patterns of the ¹⁴N and ¹⁵N peptide signals (m/z) were used for relative quantification with ProRata. <b>B</b>) Extracted ion chromatograms for the GLO1 peptide, extracted from A, showing an upregulation of GLO1 in LAB mice. No smoothing was applied.</p

Bacteria and algae diet compositions.

<p>Bacteria and algae diet compositions.</p

Anxiety-related behavior and locomotion on the elevated plus maze (EPM).

<p>High (HAB), normal (NAB) and low (LAB) anxious animals were fed with ¹⁴N or ¹⁵N enriched bacteria diets. <b>A</b>) HAB, NAB and LAB mice showed the significant (not indicated) phenotypic divergence typical of animals of the standard breeding (dotted lines and indicated in <b>B</b>) as percentage difference of the standard breeding).</p