Trophic Transfer of Macroalgal Fatty Acids in Two Urchin Species: Digestion, Egestion, and Tissue Building

Sea urchins are ecosystem engineers of nearshore benthic communities because of their influence on the abundance and distribution of macroalgal species. Urchins are notoriously inefficient in assimilation of their macroalgal diets, so their fecal production can provide a nutritional subsidy to benthic consumers that cannot capture and handle large macroalgae. We studied the assimilation of macroalgal diets by urchins by analyzing the profiles of trophic biomarkers such as fatty acids (FAs). We tracked macroalgal diet assimilation in both Strongylocentrotus droebachiensis and S. purpuratus. Juvenile S. droebachiensis and adult S. purpuratus were maintained for 180 and 70 days, respectively, on one of three monoculture diets from three algal phyla: Nereocystis luetkeana, Pyropia sp., or Ulva sp. We then analyzed FA profiles of the macroalgal tissue fed to urchins as well as urchin gonad, gut, digesta, and egesta (feces) to directly evaluate trophic modification and compare nutritional quality of urchin food sources, urchin tissues, and fecal subsidies. In the S. purpuratus assay, there were significantly more total lipids in the digesta and egesta than in the algae consumed. The FA profiles of urchin tissues differed among urchin species, all diets, and tissue types. Despite these differences, we observed similar patterns in the relationships between the urchin and macroalgal tissues for both species. Egesta produced by urchins fed each of the three diets were depleted with respect to the concentration of important long chain polyunsaturated fatty acids (LCPUFAs), but did not differ significantly from the source alga consumed. Both urchin species were shown to synthesize and selectively retain both the precursor and resulting LCPUFAs involved in the synthesis of the LCPUFAs 20:4ω6 and 20:5ω3. S. droebachiensis and S. purpuratus exhibited consistent patterns in the respective depletion and retention of precursor FAs and resulting LCPUFAs of Pyropia and Ulva tissues, suggesting species level control of macroalgal digestion or differential tissue processing by gut microbiota. For both S. droebachiensis and S. purpuratus, macroalgal diet was a surprisingly strong driver of urchin tissue fatty acids; this indicates the potential of fatty acids for future quantitative trophic estimates of urchin assimilation of algal phyla in natural settings

Nitrosopumilus maritimus gen. nov., sp. nov., Nitrosopumilus cobalaminigenes sp. nov., Nitrosopumilus oxyclinae sp. nov., and Nitrosopumilus ureiphilus sp. nov., four marine ammonia-oxidizing archaea of the phylum Thaumarchaeota

Four mesophilic, neutrophilic, and aerobic marine ammonia-oxidizing archaea, designated strains SCM1^T, HCA1^T, HCE1^T and PS0^T, were isolated from a tropical marine fish tank, dimly lit deep coastal waters, the lower euphotic zone of coastal waters, and near-surface sediment in the Puget Sound estuary, respectively. Cells are straight or slightly curved small rods, 0.15–0.26 µm in diameter and 0.50–1.59 µm in length. Motility was not observed, although strain PS0^T possesses genes associated with archaeal flagella and chemotaxis, suggesting it may be motile under some conditions. Cell membranes consist of glycerol dibiphytanyl glycerol tetraether (GDGT) lipids, with crenarchaeol as the major component. Strain SCM1^T displays a single surface layer (S-layer) with p6 symmetry, distinct from the p3-S-layer reported for the soil ammonia-oxidizing archaeon Nitrososphaera viennensis EN76^T. Respiratory quinones consist of fully saturated and monounsaturated menaquinones with 6 isoprenoid units in the side chain. Cells obtain energy from ammonia oxidation and use carbon dioxide as carbon source; addition of an α-keto acid (α-ketoglutaric acid) was necessary to sustain growth of strains HCA1^T, HCE1^T, and PS0^T. Strain PS0^T uses urea as a source of ammonia for energy production and growth. All strains synthesize vitamin B_1 (thiamine), B_2 (riboflavin), B_6 (pyridoxine), and B_(12) (cobalamin). Optimal growth occurs between 25 and 32 °C, between pH 6.8 and 7.3, and between 25 and 37 ‰ salinity. All strains have a low mol% G+C content of 33.0–34.2. Strains are related by 98 % or greater 16S rRNA gene sequence identity, sharing ~85 % 16S rRNA gene sequence identity with Nitrososphaera viennensis EN76^T. All four isolates are well separated by phenotypic and genotypic characteristics and are here assigned to distinct species within the genus Nitrosopumilus gen. nov. Isolates SCM1^T (=ATCC TSD-97^T =NCIMB 15022^T), HCA1^T (=ATCC TSD-96^T), HCE1^T(=ATCC TSD-98^T), and PS0^T (=ATCC TSD-99^T) are type strains of the species Nitrosopumilus maritimus sp. nov., Nitrosopumilus cobalaminigenessp. nov., Nitrosopumilus oxyclinae sp. nov., and Nitrosopumilus ureiphilus sp. nov., respectively. In addition, we propose the family Nitrosopumilaceae fam. nov. and the order Nitrosopumilales ord. nov. within the class Nitrososphaeria

Nitrosopumilus maritimus gen. nov., sp. nov., Nitrosopumilus cobalaminigenes sp. nov., Nitrosopumilus oxyclinae sp. nov., and Nitrosopumilus ureiphilus sp. nov., four marine ammonia-oxidizing archaea of the phylum Thaumarchaeota

Four mesophilic, neutrophilic, and aerobic marine ammonia-oxidizing archaea, designated strains SCM1^T, HCA1^T, HCE1^T and PS0^T, were isolated from a tropical marine fish tank, dimly lit deep coastal waters, the lower euphotic zone of coastal waters, and near-surface sediment in the Puget Sound estuary, respectively. Cells are straight or slightly curved small rods, 0.15–0.26 µm in diameter and 0.50–1.59 µm in length. Motility was not observed, although strain PS0^T possesses genes associated with archaeal flagella and chemotaxis, suggesting it may be motile under some conditions. Cell membranes consist of glycerol dibiphytanyl glycerol tetraether (GDGT) lipids, with crenarchaeol as the major component. Strain SCM1^T displays a single surface layer (S-layer) with p6 symmetry, distinct from the p3-S-layer reported for the soil ammonia-oxidizing archaeon Nitrososphaera viennensis EN76^T. Respiratory quinones consist of fully saturated and monounsaturated menaquinones with 6 isoprenoid units in the side chain. Cells obtain energy from ammonia oxidation and use carbon dioxide as carbon source; addition of an α-keto acid (α-ketoglutaric acid) was necessary to sustain growth of strains HCA1^T, HCE1^T, and PS0^T. Strain PS0^T uses urea as a source of ammonia for energy production and growth. All strains synthesize vitamin B_1 (thiamine), B_2 (riboflavin), B_6 (pyridoxine), and B_(12) (cobalamin). Optimal growth occurs between 25 and 32 °C, between pH 6.8 and 7.3, and between 25 and 37 ‰ salinity. All strains have a low mol% G+C content of 33.0–34.2. Strains are related by 98 % or greater 16S rRNA gene sequence identity, sharing ~85 % 16S rRNA gene sequence identity with Nitrososphaera viennensis EN76^T. All four isolates are well separated by phenotypic and genotypic characteristics and are here assigned to distinct species within the genus Nitrosopumilus gen. nov. Isolates SCM1^T (=ATCC TSD-97^T =NCIMB 15022^T), HCA1^T (=ATCC TSD-96^T), HCE1^T(=ATCC TSD-98^T), and PS0^T (=ATCC TSD-99^T) are type strains of the species Nitrosopumilus maritimus sp. nov., Nitrosopumilus cobalaminigenessp. nov., Nitrosopumilus oxyclinae sp. nov., and Nitrosopumilus ureiphilus sp. nov., respectively. In addition, we propose the family Nitrosopumilaceae fam. nov. and the order Nitrosopumilales ord. nov. within the class Nitrososphaeria

Table_11_Trophic Transfer of Macroalgal Fatty Acids in Two Urchin Species: Digestion, Egestion, and Tissue Building.docx

<p>Sea urchins are ecosystem engineers of nearshore benthic communities because of their influence on the abundance and distribution of macroalgal species. Urchins are notoriously inefficient in assimilation of their macroalgal diets, so their fecal production can provide a nutritional subsidy to benthic consumers that cannot capture and handle large macroalgae. We studied the assimilation of macroalgal diets by urchins by analyzing the profiles of trophic biomarkers such as fatty acids (FAs). We tracked macroalgal diet assimilation in both Strongylocentrotus droebachiensis and S. purpuratus. Juvenile S. droebachiensis and adult S. purpuratus were maintained for 180 and 70 days, respectively, on one of three monoculture diets from three algal phyla: Nereocystis luetkeana, Pyropia sp., or Ulva sp. We then analyzed FA profiles of the macroalgal tissue fed to urchins as well as urchin gonad, gut, digesta, and egesta (feces) to directly evaluate trophic modification and compare nutritional quality of urchin food sources, urchin tissues, and fecal subsidies. In the S. purpuratus assay, there were significantly more total lipids in the digesta and egesta than in the algae consumed. The FA profiles of urchin tissues differed among urchin species, all diets, and tissue types. Despite these differences, we observed similar patterns in the relationships between the urchin and macroalgal tissues for both species. Egesta produced by urchins fed each of the three diets were depleted with respect to the concentration of important long chain polyunsaturated fatty acids (LCPUFAs), but did not differ significantly from the source alga consumed. Both urchin species were shown to synthesize and selectively retain both the precursor and resulting LCPUFAs involved in the synthesis of the LCPUFAs 20:4ω6 and 20:5ω3. S. droebachiensis and S. purpuratus exhibited consistent patterns in the respective depletion and retention of precursor FAs and resulting LCPUFAs of Pyropia and Ulva tissues, suggesting species level control of macroalgal digestion or differential tissue processing by gut microbiota. For both S. droebachiensis and S. purpuratus, macroalgal diet was a surprisingly strong driver of urchin tissue fatty acids; this indicates the potential of fatty acids for future quantitative trophic estimates of urchin assimilation of algal phyla in natural settings.</p

Table_9_Trophic Transfer of Macroalgal Fatty Acids in Two Urchin Species: Digestion, Egestion, and Tissue Building.DOCX

<p>Sea urchins are ecosystem engineers of nearshore benthic communities because of their influence on the abundance and distribution of macroalgal species. Urchins are notoriously inefficient in assimilation of their macroalgal diets, so their fecal production can provide a nutritional subsidy to benthic consumers that cannot capture and handle large macroalgae. We studied the assimilation of macroalgal diets by urchins by analyzing the profiles of trophic biomarkers such as fatty acids (FAs). We tracked macroalgal diet assimilation in both Strongylocentrotus droebachiensis and S. purpuratus. Juvenile S. droebachiensis and adult S. purpuratus were maintained for 180 and 70 days, respectively, on one of three monoculture diets from three algal phyla: Nereocystis luetkeana, Pyropia sp., or Ulva sp. We then analyzed FA profiles of the macroalgal tissue fed to urchins as well as urchin gonad, gut, digesta, and egesta (feces) to directly evaluate trophic modification and compare nutritional quality of urchin food sources, urchin tissues, and fecal subsidies. In the S. purpuratus assay, there were significantly more total lipids in the digesta and egesta than in the algae consumed. The FA profiles of urchin tissues differed among urchin species, all diets, and tissue types. Despite these differences, we observed similar patterns in the relationships between the urchin and macroalgal tissues for both species. Egesta produced by urchins fed each of the three diets were depleted with respect to the concentration of important long chain polyunsaturated fatty acids (LCPUFAs), but did not differ significantly from the source alga consumed. Both urchin species were shown to synthesize and selectively retain both the precursor and resulting LCPUFAs involved in the synthesis of the LCPUFAs 20:4ω6 and 20:5ω3. S. droebachiensis and S. purpuratus exhibited consistent patterns in the respective depletion and retention of precursor FAs and resulting LCPUFAs of Pyropia and Ulva tissues, suggesting species level control of macroalgal digestion or differential tissue processing by gut microbiota. For both S. droebachiensis and S. purpuratus, macroalgal diet was a surprisingly strong driver of urchin tissue fatty acids; this indicates the potential of fatty acids for future quantitative trophic estimates of urchin assimilation of algal phyla in natural settings.</p

Table_7_Trophic Transfer of Macroalgal Fatty Acids in Two Urchin Species: Digestion, Egestion, and Tissue Building.DOCX

<p>Sea urchins are ecosystem engineers of nearshore benthic communities because of their influence on the abundance and distribution of macroalgal species. Urchins are notoriously inefficient in assimilation of their macroalgal diets, so their fecal production can provide a nutritional subsidy to benthic consumers that cannot capture and handle large macroalgae. We studied the assimilation of macroalgal diets by urchins by analyzing the profiles of trophic biomarkers such as fatty acids (FAs). We tracked macroalgal diet assimilation in both Strongylocentrotus droebachiensis and S. purpuratus. Juvenile S. droebachiensis and adult S. purpuratus were maintained for 180 and 70 days, respectively, on one of three monoculture diets from three algal phyla: Nereocystis luetkeana, Pyropia sp., or Ulva sp. We then analyzed FA profiles of the macroalgal tissue fed to urchins as well as urchin gonad, gut, digesta, and egesta (feces) to directly evaluate trophic modification and compare nutritional quality of urchin food sources, urchin tissues, and fecal subsidies. In the S. purpuratus assay, there were significantly more total lipids in the digesta and egesta than in the algae consumed. The FA profiles of urchin tissues differed among urchin species, all diets, and tissue types. Despite these differences, we observed similar patterns in the relationships between the urchin and macroalgal tissues for both species. Egesta produced by urchins fed each of the three diets were depleted with respect to the concentration of important long chain polyunsaturated fatty acids (LCPUFAs), but did not differ significantly from the source alga consumed. Both urchin species were shown to synthesize and selectively retain both the precursor and resulting LCPUFAs involved in the synthesis of the LCPUFAs 20:4ω6 and 20:5ω3. S. droebachiensis and S. purpuratus exhibited consistent patterns in the respective depletion and retention of precursor FAs and resulting LCPUFAs of Pyropia and Ulva tissues, suggesting species level control of macroalgal digestion or differential tissue processing by gut microbiota. For both S. droebachiensis and S. purpuratus, macroalgal diet was a surprisingly strong driver of urchin tissue fatty acids; this indicates the potential of fatty acids for future quantitative trophic estimates of urchin assimilation of algal phyla in natural settings.</p

Table_10_Trophic Transfer of Macroalgal Fatty Acids in Two Urchin Species: Digestion, Egestion, and Tissue Building.DOCX

<p>Sea urchins are ecosystem engineers of nearshore benthic communities because of their influence on the abundance and distribution of macroalgal species. Urchins are notoriously inefficient in assimilation of their macroalgal diets, so their fecal production can provide a nutritional subsidy to benthic consumers that cannot capture and handle large macroalgae. We studied the assimilation of macroalgal diets by urchins by analyzing the profiles of trophic biomarkers such as fatty acids (FAs). We tracked macroalgal diet assimilation in both Strongylocentrotus droebachiensis and S. purpuratus. Juvenile S. droebachiensis and adult S. purpuratus were maintained for 180 and 70 days, respectively, on one of three monoculture diets from three algal phyla: Nereocystis luetkeana, Pyropia sp., or Ulva sp. We then analyzed FA profiles of the macroalgal tissue fed to urchins as well as urchin gonad, gut, digesta, and egesta (feces) to directly evaluate trophic modification and compare nutritional quality of urchin food sources, urchin tissues, and fecal subsidies. In the S. purpuratus assay, there were significantly more total lipids in the digesta and egesta than in the algae consumed. The FA profiles of urchin tissues differed among urchin species, all diets, and tissue types. Despite these differences, we observed similar patterns in the relationships between the urchin and macroalgal tissues for both species. Egesta produced by urchins fed each of the three diets were depleted with respect to the concentration of important long chain polyunsaturated fatty acids (LCPUFAs), but did not differ significantly from the source alga consumed. Both urchin species were shown to synthesize and selectively retain both the precursor and resulting LCPUFAs involved in the synthesis of the LCPUFAs 20:4ω6 and 20:5ω3. S. droebachiensis and S. purpuratus exhibited consistent patterns in the respective depletion and retention of precursor FAs and resulting LCPUFAs of Pyropia and Ulva tissues, suggesting species level control of macroalgal digestion or differential tissue processing by gut microbiota. For both S. droebachiensis and S. purpuratus, macroalgal diet was a surprisingly strong driver of urchin tissue fatty acids; this indicates the potential of fatty acids for future quantitative trophic estimates of urchin assimilation of algal phyla in natural settings.</p

Table_1_Trophic Transfer of Macroalgal Fatty Acids in Two Urchin Species: Digestion, Egestion, and Tissue Building.DOCX

<p>Sea urchins are ecosystem engineers of nearshore benthic communities because of their influence on the abundance and distribution of macroalgal species. Urchins are notoriously inefficient in assimilation of their macroalgal diets, so their fecal production can provide a nutritional subsidy to benthic consumers that cannot capture and handle large macroalgae. We studied the assimilation of macroalgal diets by urchins by analyzing the profiles of trophic biomarkers such as fatty acids (FAs). We tracked macroalgal diet assimilation in both Strongylocentrotus droebachiensis and S. purpuratus. Juvenile S. droebachiensis and adult S. purpuratus were maintained for 180 and 70 days, respectively, on one of three monoculture diets from three algal phyla: Nereocystis luetkeana, Pyropia sp., or Ulva sp. We then analyzed FA profiles of the macroalgal tissue fed to urchins as well as urchin gonad, gut, digesta, and egesta (feces) to directly evaluate trophic modification and compare nutritional quality of urchin food sources, urchin tissues, and fecal subsidies. In the S. purpuratus assay, there were significantly more total lipids in the digesta and egesta than in the algae consumed. The FA profiles of urchin tissues differed among urchin species, all diets, and tissue types. Despite these differences, we observed similar patterns in the relationships between the urchin and macroalgal tissues for both species. Egesta produced by urchins fed each of the three diets were depleted with respect to the concentration of important long chain polyunsaturated fatty acids (LCPUFAs), but did not differ significantly from the source alga consumed. Both urchin species were shown to synthesize and selectively retain both the precursor and resulting LCPUFAs involved in the synthesis of the LCPUFAs 20:4ω6 and 20:5ω3. S. droebachiensis and S. purpuratus exhibited consistent patterns in the respective depletion and retention of precursor FAs and resulting LCPUFAs of Pyropia and Ulva tissues, suggesting species level control of macroalgal digestion or differential tissue processing by gut microbiota. For both S. droebachiensis and S. purpuratus, macroalgal diet was a surprisingly strong driver of urchin tissue fatty acids; this indicates the potential of fatty acids for future quantitative trophic estimates of urchin assimilation of algal phyla in natural settings.</p

Table_8_Trophic Transfer of Macroalgal Fatty Acids in Two Urchin Species: Digestion, Egestion, and Tissue Building.DOCX

<p>Sea urchins are ecosystem engineers of nearshore benthic communities because of their influence on the abundance and distribution of macroalgal species. Urchins are notoriously inefficient in assimilation of their macroalgal diets, so their fecal production can provide a nutritional subsidy to benthic consumers that cannot capture and handle large macroalgae. We studied the assimilation of macroalgal diets by urchins by analyzing the profiles of trophic biomarkers such as fatty acids (FAs). We tracked macroalgal diet assimilation in both Strongylocentrotus droebachiensis and S. purpuratus. Juvenile S. droebachiensis and adult S. purpuratus were maintained for 180 and 70 days, respectively, on one of three monoculture diets from three algal phyla: Nereocystis luetkeana, Pyropia sp., or Ulva sp. We then analyzed FA profiles of the macroalgal tissue fed to urchins as well as urchin gonad, gut, digesta, and egesta (feces) to directly evaluate trophic modification and compare nutritional quality of urchin food sources, urchin tissues, and fecal subsidies. In the S. purpuratus assay, there were significantly more total lipids in the digesta and egesta than in the algae consumed. The FA profiles of urchin tissues differed among urchin species, all diets, and tissue types. Despite these differences, we observed similar patterns in the relationships between the urchin and macroalgal tissues for both species. Egesta produced by urchins fed each of the three diets were depleted with respect to the concentration of important long chain polyunsaturated fatty acids (LCPUFAs), but did not differ significantly from the source alga consumed. Both urchin species were shown to synthesize and selectively retain both the precursor and resulting LCPUFAs involved in the synthesis of the LCPUFAs 20:4ω6 and 20:5ω3. S. droebachiensis and S. purpuratus exhibited consistent patterns in the respective depletion and retention of precursor FAs and resulting LCPUFAs of Pyropia and Ulva tissues, suggesting species level control of macroalgal digestion or differential tissue processing by gut microbiota. For both S. droebachiensis and S. purpuratus, macroalgal diet was a surprisingly strong driver of urchin tissue fatty acids; this indicates the potential of fatty acids for future quantitative trophic estimates of urchin assimilation of algal phyla in natural settings.</p

Table_5_Trophic Transfer of Macroalgal Fatty Acids in Two Urchin Species: Digestion, Egestion, and Tissue Building.DOCX

<p>Sea urchins are ecosystem engineers of nearshore benthic communities because of their influence on the abundance and distribution of macroalgal species. Urchins are notoriously inefficient in assimilation of their macroalgal diets, so their fecal production can provide a nutritional subsidy to benthic consumers that cannot capture and handle large macroalgae. We studied the assimilation of macroalgal diets by urchins by analyzing the profiles of trophic biomarkers such as fatty acids (FAs). We tracked macroalgal diet assimilation in both Strongylocentrotus droebachiensis and S. purpuratus. Juvenile S. droebachiensis and adult S. purpuratus were maintained for 180 and 70 days, respectively, on one of three monoculture diets from three algal phyla: Nereocystis luetkeana, Pyropia sp., or Ulva sp. We then analyzed FA profiles of the macroalgal tissue fed to urchins as well as urchin gonad, gut, digesta, and egesta (feces) to directly evaluate trophic modification and compare nutritional quality of urchin food sources, urchin tissues, and fecal subsidies. In the S. purpuratus assay, there were significantly more total lipids in the digesta and egesta than in the algae consumed. The FA profiles of urchin tissues differed among urchin species, all diets, and tissue types. Despite these differences, we observed similar patterns in the relationships between the urchin and macroalgal tissues for both species. Egesta produced by urchins fed each of the three diets were depleted with respect to the concentration of important long chain polyunsaturated fatty acids (LCPUFAs), but did not differ significantly from the source alga consumed. Both urchin species were shown to synthesize and selectively retain both the precursor and resulting LCPUFAs involved in the synthesis of the LCPUFAs 20:4ω6 and 20:5ω3. S. droebachiensis and S. purpuratus exhibited consistent patterns in the respective depletion and retention of precursor FAs and resulting LCPUFAs of Pyropia and Ulva tissues, suggesting species level control of macroalgal digestion or differential tissue processing by gut microbiota. For both S. droebachiensis and S. purpuratus, macroalgal diet was a surprisingly strong driver of urchin tissue fatty acids; this indicates the potential of fatty acids for future quantitative trophic estimates of urchin assimilation of algal phyla in natural settings.</p