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Supplemental Information

Figure S1

Bacterial species from 42 fish gut communities classified into the phylum level. Bar graphs for each library represent the percentage of each phylum. Scientific names of the fish hosts are listed along the horizontal axis. Habitat and food habits of these fish are described in Table S1.

DOI: 10.7287/peerj.preprints.472v2/supp-1

Figure S2

Microbiota profiles from fish guts evaluated by PCA (n = 41, k = 52, R2X = 0.154, R2Y = 0.114, Q2 = 0.059). (A) PCA score plot based on PC1 and PC2. The numbers on PCA denote the values of species diversity calculated by the Shannon diversity index. (B) PCA score plot based on PC1 and PC4. (C, D, E) PCA loading plots for PC1 (C), PC2 (D), and PC4 (E) of top 12 positives (C, D, E) and negatives (C).

DOI: 10.7287/peerj.preprints.472v2/supp-2

Figure S3

Microbiota profiles from fish guts evaluated by PCA (n = 19, k = 69, R2X = 0.175, R2Y = 0.158, Q2 = 0.018) for comparison between fish eaters and Omnivora. (A) PCA score plot based on PC1 and PC2. Symbols representing individual communities are colored by Fish eater (red) and Omnivora (green). (B) PCA loading plots for PC1.

DOI: 10.7287/peerj.preprints.472v2/supp-3

Figure S4

Supplemental Figure 4. 1H NMR spectra of methanol-soluble fraction from 14 fish feces (a) to (n). (a) to (n) fish are listed in Table S3.

DOI: 10.7287/peerj.preprints.472v2/supp-4

Figure S5

1H-13C HSQC spectra of methanol-soluble component profiles from artificial feed. Peaks were used in Figure 2. Red, amino acid; blue, unsaturated fatty acid; aqua, fatty acid and phospholipid. Peak numbers and annotated metabolites are listed in Supplementary Table S3.

DOI: 10.7287/peerj.preprints.472v2/supp-5

Figure S6

1H-13C HSQC spectra of methanol-soluble component profiles for evaluation of retention capability of water-soluble components in fish feces of Sebastiscus marmoratus. Black and red indicate the fecal samples before and after rinsing with artificial seawater (for 15 min with shaking, 3 times), respectively.

DOI: 10.7287/peerj.preprints.472v2/supp-6

Figure S7

Metabolic and microbial profiles of intestinal contents and feces of 13 species of breeding fish (Table S3). PCA scores plot for microbial profile in feces (n = 22, k = 109, R2X = 0.282, R2Y = 0.241, Q2 = 0.089), microbial profile in intestinal contents (n = 19, k = 65, R2X = 0.219, R2Y = 0.141, Q2 = 0.066), metabolic profile in feces (n = 115, k = 793, R2X = 0.224, R2Y = 0.187, Q2 = 0.138), and metabolic profile in intestinal contents (n = 20, k = 786, R2X = 0.331, R2Y = 0.152, Q2 = 0.251) are shown. Triangle indicates intestinal contents, circle indicates feces. Color indicates the feeding status of breeding fish.

DOI: 10.7287/peerj.preprints.472v2/supp-7

Table S1

List of fishes for which the sequence analysis was performed

DOI: 10.7287/peerj.preprints.472v2/supp-8

Table S2

List of methanol-soluble compounds detected in 1H-13C HSQC spectra and annotated by SpinAssign program

DOI: 10.7287/peerj.preprints.472v2/supp-9

Table S3

Supplemental Table 3. List of fishes that performed the culturing and sampling of feces

DOI: 10.7287/peerj.preprints.472v2/supp-10

Additional Information

Competing Interests

Kenji Sakata, Yasuhiro Date and Jun Kikuchi are employees of the RIKEN Center for Sustainable Resource Science (CSRS). Taiga Asakura and Seiji Yoshida are student trainees of the RIKEN CSRS.

Author Contributions

Taiga Asakura performed the experiments, analyzed the data, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Kenji Sakata performed the experiments.

Seiji Yoshida performed the experiments.

Yasuhiro Date analyzed the data, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Jun Kikuchi conceived and designed the experiments, contributed reagents/materials/analysis tools, wrote the paper, reviewed drafts of the paper.

Animal Ethics

The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):

There is no specific permission requirement for fish studies in Japan.

Field Study Permissions

The following information was supplied relating to field study approvals (i.e., approving body and any reference numbers):

There is no specific permission requirement as the sampling locations are public places. The field does not host endangered or protected species.

Grant Disclosures

The following grant information was disclosed by the authors:

This research was supported in part by Grants-in-Aid for Scientific Research (Grant No. 25513012) (to J.K.), the Advanced Low Carbon Technology Research and Developmental Program (Grant No. 200210023, ALCA to J.K.) from the Ministry of Education, Culture and Sports, and RIKEN Junior Research Associate Program.

Funding

This research was supported in part by Grants-in-Aid for Scientific Research (Grant No. 25513012) (to J.K.), the Advanced Low Carbon Technology Research and Developmental Program (Grant No. 200210023, ALCA to J.K.) from the Ministry of Education, Culture and Sports, and the RIKEN Junior Research Associate Program. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.


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