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Appendix
Appendix
241
Appendix I
Description of collection sites of different macroalgae.
Species Collection site Location CHLOROPHYTA Ulvales Ulva lactuca Veraval N 20º 54.87'; E 70º 20.83' Ulva fasciata Veraval N 20º 54.87'; E 70º 20.83' Ulva taeniata Veraval N 20º 54.87'; E 70º 20.83' Ulva pertusa Kalubhar island N 22º 26.21'; E 69º 35.12’ Ulva reticulata Kalubhar island N 22º 26.21'; E 69º 35.12’ Ulva beytensis Kalubhar island N 22º 26.21'; E 69º 35.12’ Ulva compressa Kalubhar island N 22º 26.21'; E 69º 35.12’ Ulva rigida Veraval N 20º 54.87'; E 70º 20.83' Ulva linza Veraval N 20º 54.87'; E 70º 20.83' Ulva flexuosa Veraval N 20º 54.87'; E 70º 20.83' Ulva erecta Kalubhar island N 22º 26.21'; E 69º 35.12’ Ulva prolifera Kalubhar island N 22º 26.21'; E 69º 35.12’ Bryopsidales Caulerpa scalpelliformis Veraval N 20º 54.87'; E 70º 20.83' Caulerpa veravalensis Veraval N 20º 54.87'; E 70º 20.83' Caulerpa racemosa Veraval N 20º 54.87'; E 70º 20.83' Caulerpa racemosa v. corynephora Okha N 22º 27.06'; E 69º 4.02' Caulerpa racemosa v. occidentalis Okha N 22º 28.50'; E 69º 4.54' Caulerpa microphysa Veraval N 22º 28.43'; E 69º 4.17' Caulerpa verticillata Okha N 22º 28.39'; E 69º 4.50' Caulerpa sertularioides Okha N 22º 28.42'; E 69º 4.51' Codium dwarkense Veraval N 20º 54.87'; E 70º 20.83' Bryopsis pennata Okha N 22º 28.40'; E 69º 4.50' Bryopsis plumosa Okha N 22º 28.44'; E 69º 4.50' Trichosolen mucronatus Okha N 22º 28.41'; E 69º 3.58' Udotea indica Okha N 22º 28.43'; E 69º 4.10' Halimeda discoides Veraval N 20º 54.87'; E 70º 20.83' Halimeda tuna Veraval N 20º 54.87'; E 70º 20.83' Ulotrichales Monostroma oxyspermum Achara N 16º 11.59’; E 73º 26.38’ Siphonocladales Chamaedoris auriculata Veraval N 20º 54.87'; E 70º 20.83' Cladophoropsis javanica Okha N 22º 28.44'; E 69º 4.50' Valoniopsis pachynema Veraval N 20º 54.87'; E 70º 20.83' Cladophorales Chaetomorpha linum Kalubhar island N 22º 26.21'; E 69º 35.12’ Acrosiphonia orientalis Okha N 22º 28.46'; E 69º 04.34' PHAEOPHYTA Dictyotales Padina tetrastomatica Veraval N 20º 54.87'; E 70º 20.83' Padina gymnospora Kalubhar island N 22º 26.21'; E 69º 35.12’
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Dictyopteris deliculata Kalubhar island N 22º 26.21'; E 69º 35.12’ Dictyota pinnatifida Kalubhar island N 22º 26.21'; E 69º 35.12’ Dictyota bartayresiana Kalubhar island N 22º 26.21'; E 69º 35.12’ Dictyota dichotoma Kalubhar island N 22º 26.21'; E 69º 35.12’ Dictyota cervicornis Kalubhar island N 22º 26.21'; E 69º 35.12’ Dictyota ciliolata Kalubhar island N 22º 26.21'; E 69º 35.12’ Dictyota haukiana Kalubhar island N 22º 26.21'; E 69º 35.12’ Stoechospermum marginatum Veraval N 20º 54.87'; E 70º 20.83' Lobophora variegata Veraval N 20º 54.87'; E 70º 20.83' Spatoglossum asperum Veraval N 20º 54.87'; E 70º 20.83' Fucales Sargassum tenerrimum Veraval N 20º 54.87'; E 70º 20.83' Sargassum johnstonii Okha N 22º 28.54'; E 69º 04.59' Sargassum sp. Kalubhar island N 22º 26.21'; E 69º 35.12’ Sargassum carpophyllum Kalubhar island N 22º 26.21'; E 69º 35.12’ Sargassum plagiophyllum Shivrajpur N 22º 19.87'; E 68º 56.95_ Sargassum cinereum Veraval N 20º 54.87'; E 70º 20.83' Sargassum cinctum Porbandar N 21º 38.24'; E 69º 35.81' Hormophysa cuneiformis Okha N 22º 28.43'; E 69º 04.10' Cystoseira indica Veraval N 20º 54.87'; E 70º 20.83' Cystoseira trinodis Dhani island N 22º 24.81’; E 69º 32.24’ Ectocarpales Hincksia mitchelliae Dhani island N 22º 24.81’; E 69º 32.24’ Scytosiphon lomentaria Dhani island N 22º 24.81’; E 69º 32.24’ RHODOPHYTA Gracilariales Gracilaria dura Adri N 20º 57.58'; E 70º 16.76' Gracilaria salicornia Veraval N 20º 54.87'; E 70º 20.83' Gracilaria textorii Veraval N 20º 54.87'; E 70º 20.83' Gracilaria corticata Veraval N 20º 54.87'; E 70º 20.83' Gracilaria corticata v. cylindrica Veraval N 20º 54.87'; E 70º 20.83' Gracilaria corticata v. folifera Veraval N 20º 54.87'; E 70º 20.83' Gracilaria debilis Okha N 22º 28.44'; E 69º 03.58' Gracilaria verrucosa Okha N22º 26.38'; E 69º 03.30' Gigartinales Sarconema scinaioides Veraval N 20º 54.87'; E 70º 20.83' Sarconema filiforme Veraval N 20º 54.87'; E 70º 20.83' Hypnea valentiae Okha N 22º 28.54'; E 69º 04.38' Hypnea musciformis Okha N 22º 28.51'; E 69º 04.48' Hypnea spinella Veraval N 20º 54.87'; E 70º 20.83' Solieria robusta Kalubhar island N 22º 26.21'; E 69º 35.12’ Bangiales Pyropia tenera Malvan N 16 º 03. 59'; E 73º 27. 31’ Pyropia yoezensis Malvan N 16 º 03. 59'; E 73º 27. 31’ Pyropia acanthophora Malvan N 16 º 03. 59'; E 73º 27. 31’ Pyropia acanthophora v. brasilensis Malvan N 16 º 03. 59'; E 73º 27. 31’ Pyropia sp. Malvan N 16 º 03. 59'; E 73º 27. 31’
Appendix
243
Rhodymeniales Rhodymenia sonderi Veraval N 20º 54.87'; E 70º 20.83' Coelarthrum muelleri Okha N 22º 28.47'; E 69º 04.55' Botryocladia leptopoda Okha N 22º 28.49'; E 69º 04.56' Botryocladia botryoides Boria reef N 22 º 24.49'; E 69º 13.33’ Gastroclonium iyengarii Dhani island N 22º 24.81’; E 69º 32.24’ Champia parvula Dhani island N 22º 24.81’; E 69º 32.24’ Gelidiopsis variabilis Okha N 22º 28.52'; E 69º 04.47' Gelidiales Gelidiella acerosa Veraval N 20º 54.87'; E 70º 20.83' Halymeniales Cryptonemia undulata Okha N 22º 28.48'; E 69º 04.56' Halymenia porphyraeformis Okha N 22º 28.51'; E 69º 04.50' Grateloupia indica Okha N 22º 28.50'; E 69º 04.55' Grateloupia filicina Okha N 22º 28.43'; E 69º 04.10' Ceramiales Odonthalia veravalensis Veraval N 20º 54.87'; E 70º 20.83' Acanthophora specifera Okha N 22º 28.39'; E 69º 03.56' Acanthophora nayadiformis Okha N 22º 28.48'; E 69º 04.37' Laurencia cruciata Veraval N 20º 54.87'; E 70º 20.83' Laurencia obstusa Veraval N 20º 54.87'; E 70º 20.83' Laurencia papillosa Veraval N 20º 54.87'; E 70º 20.83' Laurencia majusculus Veraval N 20º 54.87'; E 70º 20.83' Laurencia sp. Veraval N 20º 54.87'; E 70º 20.83' Polysiphonia ferulacea Veraval N 20º 54.87'; E 70º 20.83' Griffithsia corallinoides Okha N 22º 28.44'; E 69º 04.49' Corallinales Jania rubens Okha N 22º 28.50'; E 69º 04.33' Scinaia monoliformis Okha N 22º 28.48'; E 69º 04.55'
Appendix
244
Appendix II
Map showing the sampling locations along the Gujarat coast
Appendix
245
Appendix III
Fatty acid composition of different macroalgal species given in % of total fatty acid methyl esters (TFAs), expressed as means ± SD (n=3).
FAs 1 2 3 4 5 6 7 8 9 10 11 12 C12:0 1.1±0.1 0.2±0.1 0.7±0.2 0.5±0.2 0.6±0.1 1.1±0.9 0.5±0.2 0.6±0.4 0.6±0.3 1.7±0.6 0.8±0.1 0.8±0.4 C14:0 3.3±0.5 0.7±0.1 1.8±0.3 2.6±0.3 3.8±0.4 2.6±0.6 2.1±0.2 3.7±4.1 2.1±0.1 3.4±0.2 1.7±0.1 2.1±0.4 C15:0 1.0±0.1 0.3±0.2 0.5±0.1 0.5±0.1 0.2±0.01 0.8±0.1 0.6±0.2 0.4±0.1 0.7±0.3 1.0±0.1 1.1±0.2 0.7±0.2 C16:0 39.3±1.0 25.1±1.6 34.2±5.2 25.2±1.7 34±3.0 23.3±1.5 29.4±0.5 28.2±1.6 30.6±8.3 26.2±0.8 31±0.8 29.6±0.3 C17:0 0.5±0.1 0.2±0.1 0.3±0.1 0.2±0.01 0.2±0.02 0.5±0.2 0.3±0.1 0.2±0.1 0.1±0.1 0.5±0.1 0.2±0.1 0.4±0.3 C18:0 10.2±2.7 1.6±0.3 5.3±0.8 4.7±0.3 7.9±2.0 2.5±0.4 2.6±0.1 2.4±1.1 3.1±1.6 3.3±0.2 2.1±0.1 2.7±0.6 C20:0 0.6±0.1 0.2±0.1 2.4±2.4 0.3±0.01 0.7±0.2 0.3±0.1 0.6±0.1 0.5±0.2 0.8±0.6 0.3±0.4 1.0±0.5 1.0±0.3 C22:0 3.4±0.1 1.2±0.2 1.6±0.2 1.1±0.2 1.3±0.3 0.9±0.1 1.1±0.1 1.2±1.3 0.4±0.2 0.6±0.1 1.2±0.1 1.4±0.5 C24:0 0.5±0.1 n. d. n. d. 0.2±0.01 n. d. n. d. 0.3±0.1 0.5±0.3 n. d. 0.3±0.1 n. d. 0.6±0.5 C16:1(n-7) 2.5±0.4 1.9±0.3 1.8±0.3 5.2±0.1 2.9±0.3 1.3±0.3 1.4±0.2 3.4±2.7 n. d. 5.8±0.5 n. d. 1.5±0.3C16:1(n-9) 3.4±1.0 1.9±0.8 1.2±0.5 n. d. n. d. 2.1±0.3 3.6±0.1 1.9±1.8 1.5±0.4 0.0 3.1±2. 4.8±0.3 C17:1(n-7) 0.7±0.1 0.2±0.1 0.0 0.6±0.3 0.2±0.01 0.3±0.1 0.3±0.1 0.3±0.3 0.1±0.1 0.4±0.1 0.4±0.1 0.5±0.1 C18:1(n-9) 3.8±0.3 3.3±0.6 2.4±0.6 3.6± 4.0±0.3 6.4±1.1 2.6±0.1 2.4±0.6 2.4±0.4 1.3±0.1 2.4±0.4 2.7±0.3 C18:1(n-9) trans
n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d.
C20:1(n-9) 0.4±0.1 0.3±0.1 n. d. n. d. n. d. 0.1±0.01 n. d. n. d. 0.1±0.1 n. d. n. d. n. d. C22:1(n-9 1.4±0.1 n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. C18:2(n-6) 5.2±0.7 n. d. 8.1±0.3 7.6±0.6 7.3±2.4 n. d. 10.6±0.1 6.0±4.2 n. d. 10.8±0.3 13.4±0.2 14.3±0.5 C18:3(n-6) 0.5±0.2 1.0±0.1 2.9±0.8 0.5±0.1 1.0±0.3 0.4±0.1 1.3±0.3 0.5±0.3 0.7±0.2 1.0±0.1 1.0±0.2 1.2±0.3 C18:3(n-3) 14.3±0.2 38.9±1.2 22±2.4 22.8±1.0 17.3±1.7 35.5±2.0 27.5±0.2 23.2±3.5 28.5±5.0 36.8±0.6 29.2±0.6 24±2.4 C18:4(n-3) 2.7±0.4 9.5±0.6 7.5±1.5 19.9±0.8 14.7±1.2 11.2±1.7 8.4±0.7 20±11 15.5±2.5 3.6±0.2 4.8±0.2 5.4±1.7C20:3(n6) 0.4±0.1 0.4±0.1 1.2±0.6 0.3±0.1 n. d. 0.3±0.2 0.9±0.1 0.2±0.2 0.4±0.2 0.6±0.1 0.7±0.1 0.9±0.3 C20:3(n3) 0.6±0.1 0.3±0.3 0.8±0.2 1.0±0.1 n. d. 0.3±0.1 0.7±0.2 0.9±0.1 0.8±0.5 n. d. 0.5±0.2 0.8±0.5 C20:4(n-6) 0.9±0.1 1.3±0.2 2.0±1.4 0.5±0.1 0.9±0.3 1.7±1.0 1.8±0.1 1.0±0.8 1.4±0.5 n. d. 1.4±0.2 1.8±0.3 C20:5(n-3) 0.9±0.7 0.6±0.1 1.7±1.0 0.5±0.1 1.0±0.2 0.7±0.1 1.4±0.2 0.8±0.7 1.9±0.6 0.5±0.1 1.2±0.1 1.2±0.1 C22:6(n-3) 2.5±0.4 3.4±0.3 1.9±0.3 2.1±0.2 2.6±0.4 3.1±0.5 1.4±0.3 1.6±0.5 1.5±1.0 0.7±0.1 1.3±0.3 1.9±1.4
n.d.- not detected.
Table continued..
Appendix
246
FAs 13 14 15 16 17 18 19 20 21 22 23 24 C12:0 0.3±0.1 0.1±0.01 0.2±0.1 0.5±0.3 1.1±0.2 1.7±0.1 0.4±0.3 0.5±0.3 0.6±0.1 0.0 0.9±0.2 0.3±0.1 C14:0 2.4±0.2 0.8±0.1 1.2±0.2 4.4±3.0 4.4±1.0 3.6±0.8 7.0±3.5 2.8±0.3 11.1±1.1 3.4±2.8 5.6±3.1 7.0±1.9 C15:0 0.2±0.01 0.0 0.1±0.01 0.9±0.2 0.5±0.1 0.7±0.4 0.2±0.01 0.5±0.4 0.3±0.01 0.8±0.3 0.6±0.3 0.4±0.1 C16:0 30.1±0.6 25.2±0.9 30.7±3.5 44.6±1.7 39.4±3.5 27.2±1.3 29.5±2.9 34.4±0.6 43±4.4 32.3±3.5 36.6±5.3 36.9±8.1 C17:0 0.4±0.1 0.1±0.01 0.1±0.01 0.6±0.2 1.0±0.6 0.6±0.1 0.3±0.1 n. d. 0.7±0.1 n. d. n. d. 0.3±0.1 C18:0 2.0±0.2 0.8±0.3 1.1±0.4 3.1±0.3 3.2±0.5 3.0±0.4 2.3±0.4 n. d. 7.0±0.6 2.0±0.4 3.2±0.9 2.7±1.0C20:0 0.2±0.1 n.d. n. d. 0.5±0.3 n. d. 0.3±0.1 0.3±0.1 n. d. 0.7±0.1 0.8±0.1 0.7±0.6 0.3±0.2 C22:0 0.4±0.1 n.d. n. d. 0.3±0.2 n. d. 1.1±0.3 0.4±0.2 n. d. 1.2±0.1 1.1±0.3 1.0±0.2 0.5±0.2 C24:0 3.5±0.1 n. d. n. d. 3.3±3.2 3.6±1.0 2.2±1.1 3.6±1.6 1.3±0.8 0.9±0.1 1.5±0.5 2.9±1.1 0.9±0.6 C16:1(n-7) 1.9±0.1 1.4±0.2 2.9±0.5 2.1±0.3 3.5±3.8 3.2±0.5 3.3±3.5 1.9±0.6 1.8±0.1 1.8±0.7 2.2±0.6 1.9±0.5 C16:1(n-9) 2.5±0.1 0.1±0.01 n. d. n. d. 5.6±1.1 n. d. n. d. n. d. n. d. 3.4±1.7 4.6±0.7 0.5±0.8 C17:1(n-7) 0.3±0.1 0.3±0.02 0.1±0.01 0.4±0.1 0.0 n. d. 0.2±0.2 n. d. n. d. n. d. n. d. 0.2±0.2 C18:1(n-9) 1.8±0.1 0.3±0.01 2.6±0.8 0.4±0.2 1.0±0.1 0.5±0.2 0.7±0.4 0.3±0.1 4.8±0.1 1.3±0.3 1.4±0.5 4.1±2.8 C18:1(n-9) trans
n. d. n. d. n. d. n. d. n. d. n. d. 0.6±0.4 n. d. n. d. n. d. n. d. 1.3±1.2
C20:1(n-9) n. d. n. d. 0.2±0.1 n. d. n. d. n. d. 0.4±0.2 n. d. n. d. n. d. n. d. n. d. C22:1(n-9 n. d. n. d. n. d. n. d. n. d. 1.5±0.1 0.5±0.1 n. d. n. d. n. d. n. d. 0.2±0.2C18:2(n-6) 8.8±0.3 7.1±0.3 5.5±0.6 6.8±2.2 11.1±1.4 12.2±0.8 7.4±3.3 13.5±0.7 8.9±0.7 8.2±2.8 5.2±2.4 12.4±0.8 C18:3(n-6) 1.1±0.1 1.0±0.1 0.4±0.1 0.7±0.5 1.2±0.7 1.8±0.5 4.3±0.5 1.4±0.4 1.6±0.1 1.9±0.3 3.0±0.5 1.3±0.1 C18:3(n-3) 29.8±0.9 48±0.9 45.5±0.9 15.7±5.9 14.7±4.0 26.1±2.4 15.8±6.3 28.2±1.8 10.7±1.6 28.3±7.7 21.8±4.4 10.8±1 C18:4(n-3) 1.3±0.1 1.7±0.2 0.6±0.1 4.5±3.3 2.4±0.7 2.2±1.0 5.5±2.3 2.2±0.5 1.0±0.2 3.3±0.5 2.5±2.9 1.6±0.9 C20:3(n6) 0.8±0.1 0.5±0.1 0.4±0.1 1.0±0.5 n. d. 1.1±0.1 2.0±0.3 0.9±0.5 0.8±0.1 1.3±0.9 0.7±0.3 2.0±0.4 C20:3(n3) 0.3±0.1 0.2±0.1 0.1±0.01 n. d. n. d. n. d. 0.7±0.2 1.1±0.1 n. d. n. d. n. d. 0.8±0.7 C20:4(n-6) 4.6±0.1 7.8±0.1 2.2±0.4 3.0±1.3 3.2±0.1 5.0±0.6 9.8±4.1 2.0±0.8 3.0±0.1 6.6±1.8 4.4±0.9 7.5±0.4 C20:5(n-3) 5.4±0.1 3.6±0.1 5.1±0.8 3.6±0.2 3.9±0.5 4.7±1.5 3.6±0.7 4.7±0.9 0.7±0.2 1.8±0.4 2.8±0.9 5.7±0.4 C22:6(n-3) 0.8±0.1 0.6±0.1 0.4±0.3 1.1±0.3 n. d. 1.4±0.2 0.7±0.3 1.8±0.9 1.3±0.1 n. d. n. d. n. d. SFA 39.6±1.0 27.1±1.0 33.4±3.1 58.2±6.4 n. d. 40.3±2.0 44±2.7 41.4±0.9 65.4±2.6 42.1±6.2 51.5±4.4 49.3±10.8 MUFA 6.4±0.1 1.9±0.2 5.7±1.4 2.9±0.4 10.2±5 5.2±0.3 6.1±3.0 2.2±0.6 6.6±0.1 6.5±2.6 8.2±1.1 8.7±0.2 PUFA 54±1.0 71.1±0.9 60.8±3.0 39.1±6.0 36.5±3 54.6±1.9 50±5.3 56.8±1.1 28±2.6 51.5±7.9 40.4±3.4 42.3±10.5 n6/n3 0.4±0.1 0.3±0.01 0.2±0.01 0.6±0.1 0.8±0.3 0.6±0.01 1.0±0.3 0.5±0.1 1.1±0.1 0.6±0.2 0.5±0.1 1.9±1.6 U.I. 177.5±3 226±2.6 196.6±11 128.3±18 122.1±7 176.7±6.1 172.6±6 176.9±3 91±7.8 166±26 137±11.6 144.3±33 AI 0.5±0.02 0.4±0.02 0.5±0.08 1.2±0.3 1.0±0.13 0.5±0.05 0.7±0.05 0.6±0.02 1.6±0.22 0.6±0.17 0.9±0.12 0.9±0.41 TI 0.3±0.01 0.2±0.01 0.2±0.02 0.6±0.12 0.7±0.1 0.3±0.03 0.4±0.11 0.3±0.02 1.3±0.23 0.4±0.15 0.5±0.06 0.9±0.8
n.d.- not detected.
Table continued..
Appendix
247
FAs 25 26 27 28 29 30 31 32 33 34 35 36 C12:0 0.4±0.2 1.0±0.4 0.5±0.1 1.7±0.3 0.3±0.1 1.5±0.9 1.1±0.5 0.6±0.3 1.3±0.4 0.9±0.2 0.4±0.1 0.3±0.2 C14:0 2.8±0.7 6.8±0.5 6.2±0.1 5.7±0.4 7.7±0.7 7.9±0.7 8.5±2.9 6.8±1.0 5.8±1.3 7.0±0.3 7.5±0.5 11.2±1.3 C15:0 0.5±0.3 0.9±0.3 0.5±0.1 0.8±0.1 0.4±0.1 1.4±0.7 1.0±0.7 0.7±0.5 0.6±0.3 0.7±0.1 0.6±0.1 1.4±1.7 C16:0 35±2.2 30±3.8 29.6±0.8 26.7±3.9 33.5±3.3 29.3±4.5 31.6±1.4 24.9±2.2 30.6±3.7 32.9±0.6 20.2±0.5 32.2±2.7 C17:0 0.3±0.1 0.5±0.1 0.4±0.1 0.6±0.1 0.3±0.1 1.4±1.8 0.3±0.2 0.9±0.4 0.7±0.4 0.3±0.1 0.4±0.1 0.5±0.4 C18:0 1.2±0.2 2.8±08 2.0±0.4 17.4±7.0 1.3±0.9 2.4±0.8 2.1±0.5 2.2±0.7 1.2±0.2 3.7±0.3 2.5±0.1 4.0±2.2C20:0 0.3±0.1 n. d. 0.1±0.01 1.1±0.3 0.2±0.1 0.4±0.1 0.2±0.1 n. d. n. d. 0.3±0.1 1.8±1.0 0.4±0.1 C22:0 0.6±0.4 n. d. 0.2±0.01 1.1±0.1 0.2±0.1 0.5±0.1 0.5±0.3 0.4±0.2 0.6±0.2 0.4±0.1 0.3±0.1 0.2±0.2 C24:0 2.5±0.6 3.6±1.6 2.5±0.9 0.0 0.0 0.9±0.6 1.0±0.4 0.9±0.5 0.9±0.4 0.3±0.1 0.1±0.1 0.2±0.2 C16:1(n-7) 2.3±2.0 2.3±0.6 2.3±0.2 1.0±0.7 5.8±1.5 1.7±0.4 9.2±6.5 1.7±0.7 1.9±0.2 2.7±0.2 4.8±0.3 1.0±0.5 C16:1(n-9) n. d. n. d. n. d. n. d. 2.9±0.6 3.7±2.0 1.8±0.2 2.0±0.4 3.7±0.5 1.8±0.2 5.3±0.4 1.6±0.4 C17:1(n-7) n. d. n. d. n. d. n. d. 0.2±0.1 n. d. n. d. n. d. 0.2±0.1 0.5±0.1 n. d. n. d. C18:1(n-9) 1.3±0.3 2.0±1.5 1.0±0.1 1.9±1.6 4.1±0.6 3.4±0.6 3.7±0.8 16.7±0.1 2.9±0.5 6.0±0.4 5.3±0.3 6.9±0.6 C18:1(n-9) trans
n. d. n. d. 1.7±0.7 n. d. n. d. n. d. n. d. 1.5±0.6 n. d. 3.5±0.1 1.5±1.1 1.6±0.6
C20:1(n-9) 1.3±0.7 n. d. 1.4±0.4 0.5±0.2 0.6±0.4 0.0 0.8±0.4 n. d. n. d. n. d. n. d. n. d. C22:1(n-9 0.7±0.4 n. d. 1.1±0.4 n. d. n. d. 0.3±0.1 0.0 n. d. n. d. 0.4±0.2 n. d. n. d.C18:2(n-6) 7.1±1.8 12.8±5.0 11.4±0.3 4.9±1.9 4.6±0.4 12.3±0.6 8.6±0.7 8.0±2.0 17.2±3.8 8.3±1.0 7.6±0.6 9.2±1.2 C18:3(n-6) 0.8±0.2 1.1±0.4 0.9±0.1 1.4±0.4 3.0±0.4 8.9±1.7 9.3±4.1 6.1±3.5 2.5±0.4 2.2±0.2 2.8±0.2 1.1±0.4 C18:3(n-3) 19.8±8.9 11.6±3.2 6.8±1.5 25.3±2.4 16.1±2 9.0±3.8 6.5±0.7 6.4±3.5 16.1±7.6 n. d. n. d. n. d. C18:4(n-3) 2.9±0.6 1.8±0.8 1.6±0.3 3.6±0.3 0.7±0.4 1.8±1.3 1.4±0.3 6.8±3.4 0.9±0.5 n. d. 23.5±0.6 9.4±0.4 C20:3(n6) 0.9±0.4 0.7±0.3 0.7±0.1 n. d. 0.3±0.1 1.0±0.7 1.2±0.7 1.5±0.5 0.6±0.1 1.2±0.1 1.7±0.1 1.5±1.3 C20:3(n3) 1.0±0.5 0.8±0.3 0.9±0.3 n. d. 0.2±0.1 0.7±0.7 0.4±0.2 1.0±0.4 n. d. 1.2±0.1 1.9±0.3 0.5±0.5 C20:4(n-6) 1.1±0.3 3.7±1.1 7.9±1.2 0.8±0.5 10.7±0.8 5.8±1.4 4.1±0.5 5.0±0.2 n. d. 9.2±0.8 10.6±0.6 15.4±1.3 C20:5(n-3) 11±4 9.3±3.2 11.4±1.0 1.3±0.8 6.0±0.4 2.9±0.5 2.7±0.5 2.1±1.2 3.4±1.4 1.2±0.1 1.2±0.2 1.5±0.3 C22:6(n-3) 5.1±2.1 5.9±1.9 7.1±0.9 1.2±0.8 0.9±0.4 2.3±0.9 3.5±1.7 3.6±0.8 1.9±0.6 n. d. n. d. n. d. n.d.- not detected.
Table continued..
Appendix
248
FAs 37 38 39 40 41 42 43 44 45 46 47 48 C12:0 1.4±0.9 0.6±0.3 0.6±0.1 n. d. 0.3±0.1 0.4±0.2 0.4±0.2 0.6±0.2 0.2±0.1 0.5±0.1 1.1±0.4 0.6±0.1 C14:0 4.6±0.3 5.1±1.0 8.2±1.0 7.4±1.0 14.9±0.5 10.7±1.2 24.7±4.0 14.1±1.2 12.3±0.2 7.5±0.7 5.4±0.2 5.5±0.7 C15:0 0.8±0.1 0.7±0.5 0.9±0.1 2.1±0.6 0.7±0.1 0.4±0.1 0.9±0.2 0.8±0.1 0.7±0.2 0.7±0.1 1.9±2.2 0.7±0.1 C16:0 41.5±4.9 18.8±1.3 24.5±9.8 18.4±2.0 20.1±0.4 19±1.2 27.7±1.2 28.9±2.1 35±3.0 36.9±1.0 31.7±4.1 33.7±0.4 C17:0 0.6±0.2 0.5±0.1 0.6±0.4 0.5±0.1 0.3±0.1 0.1±0.01 0.5±0.2 0.3±0.1 0.1±0.1 0.3±0.1 0.4±0.1 0.4±0.1 C18:0 4.5±0.8 1.8±0.7 2.6±0.4 4.8±0.1 2.7±1.1 2.1±0.4 3.5±3.1 1.5±0.2 2.1±0.2 2.2±1.7 4.0±0.2 6.6±0.8 C20:0 0.4±0.1 0.6±0.3 0.8±0.5 1.4±0.3 0.3±0.1 1.0±0.2 0.7±0.6 0.8±0.1 0.4±0.1 0.4±0.01 0.4±0.2 0.7±0.2 C22:0 1.0±0.2 0.3±0.1 0.4±0.1 1.3±0.3 0.4±0.1 0.3±0.1 0.5±0.1 0.5±0.2 0.1±0.1 0.6±0.01 0.8±0.3 0.7±0.1 C24:0 1.0±0.6 0.5±0.2 0.4±0.1 0.8±0.1 0.2±0.1 0.2±0.01 0.5±0.1 0.7±0.2 0.2±0.01 0.4±0.01 0.6±0.2 0.6±0.1 C16:1(n-7) 0.7±0.2 2.8±0.5 2.1±0.5 2.5±0.5 0.7±0.2 10.1±0.5 1.7±0.5 1.0±0.3 8.3±0.1 5.3±0.4 3.2±0.5 7.4±0.6 C16:1(n-9) 2.8±0.7 2.0±0.4 4.2±3.6 2.0±0.2 1.7±0.1 1.8±0.3 1.8±1.2 1.5±0.4 4.1±6.5 0.0 0.0 0.0 C17:1(n-7) 0.8±0.3 0.4±0.5 0.6±0.4 0.0 0.2±0.1 0.1±0.1 0.6±0.3 0.3±0.1 0.4±0.2 0.3±0.2 0.4±0.1 0.3±0.2 C18:1(n-9) 3.5±0.2 5.6±1.4 7.5±3.3 5.7±0.4 6.4±0.4 6.4±0.3 11.4±0.3 9.2±1.1 12.6±0.3 7.1±0.1 7.1±0.8 6.8±0.3 C18:1(n-9) trans
3.5±0.5 2.3±0.9 2.0±0.7 1.8±0.1 2.8±0.1 3.3±0.8 n. d. n. d. n. d. 1.4±0.4 1.4±1.1 1.6±0.2
C20:1(n-9) n. d. n. d. n. d. n. d. n. d. n. d. 0.8±0.1 0.3±0.2 n. d. 1.9±0.2 1.9±0.1 1.0±0.2 C22:1(n-9 n. d. n. d. n. d. n. d. 0.1±0.1 0.3±0.5 0.7±0.3 0.3±0.1 n. d. 1.7±0.1 1.1±0.1 1.2±0.1 C18:2(n-6) 4.3±0.1 3.0±0.3 3.8±1.5 6.3±0.4 5.3±0.4 0.7±0.4 3.2±1.7 2.4±0.7 0.6±0.3 4.0±0.3 3.3±0.3 5.0±0.4 C18:3(n-6) 0.7±0.5 1.7±0.8 1.6±0.4 3.6±0.5 1.3±0.1 2.0±0.1 3.7±3.0 4.0±0.2 0.4±0.2 n. d. 0.5±0.1 n. d. C18:3(n-3) n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d.C18:4(n-3) 3.3±1.3 18.7±2.0 13.4±6.0 15.9±1.5 17.4±0.4 22.4±0.6 0.0 3.6±0.7 4.8±0.5 4.5±0.5 8.2±2.3 5.4±0.5 C20:3(n6) 1.5±0.3 2.8±0.7 2.8±0.4 1.8±0.3 2.8±0.1 0.3±0.1 3.6±1.4 0.9±0.1 1.3±0.3 0.6±0.1 0.7±0.1 0.9±0.1 C20:3(n3) 1.7±0.6 5.2±1.1 3.4±2.5 2.3±0.2 8.0±0.3 1.7±1.2 3.9±3.0 1.4±0.2 2.5±0.4 1.0±0.1 1.2±0.5 0.7±0.1 C20:4(n-6) 19.0±1.0 14.5±1.1 10.9±2.1 12.2±0.8 9.7±0.2 9.0±0.2 6.2±3.3 10.2±1.4 11.2±1.3 18.1±0.6 18.7±0.7 15.6±0.3 C20:5(n-3) 2.4±0.9 12.2±0.2 8.4±3.7 9.4±0.2 2.8±0.3 7.2±0.1 2.6±0.2 16.7±1.2 2.9±0.5 3.5±0.4 5.9±1.5 3.8±0.3 C22:6(n-3) n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d.n.d.- not detected.
Table continued..
Appendix
249
FAs 49 50 51 52 53 54 55 56 57 58 59 60 C12:0 0.8±0.1 0.7±0.1 0.7±0.2 0.3±0.1 0.3±0.1 0.8±0.1 0.7±0.2 0.2±0.1 0.0 0.1±0.01 1.3±0.1 1.0±0.5 C14:0 3.7±0.5 6.6±0.3 4.6±1.7 5.4±1.3 5.0±1.2 7.1±0.3 6.3±0.2 7.7±0.8 8.7±0.3 1.9±0.1 4.2±0.1 5.2±0.7 C15:0 0.8±0.6 0.7±0.1 0.6±0.1 0.4±0.1 0.4±0.1 0.5±0.1 0.8±0.1 0.4±0.01 2.1±2.2 0.4±0.2 2.1±0.1 0.9±0.3 C16:0 37.7±3.9 28.9±1.7 35.4±4.3 26±17.3 26.4±0.5 34.5±6.0 26.8±1.8 23.5±1.3 41.5±3.1 27.8±1.5 58.1±1.5 44.8±2.7 C17:0 0.3±0.2 0.3±0.1 0.3±0.1 0.2±0.1 0.4±0.2 0.6±0.8 0.4±0.3 0.2±0.01 0.4±0.2 0.2±0.01 1.9±0.1 1.1±0.6 C18:0 7.0±5.1 4.3±0.3 0.7±0.5 2.3±0.6 6.6±2.1 1.8±0.1 2.6±0.5 1.5±0.4 3.6±0.5 2.6±0.1 5.6±0.2 4.8±3.3 C20:0 0.6±0.3 0.4±0.1 0.2±0.1 0.2±0.1 0.4±0.01 0.4±02 0.3±0.1 0.6±0.1 1.4±0.7 0.2±0.2 1.1±0.1 0.7±0.4 C22:0 1.0±0.6 0.4±0.1 0.5±0.1 0.6±0.2 0.7±0.01 0.9±0.4 0.6±0.2 1.4±0.6 0.8±0.2 n. d. 1.0±0.8 1.0±0.5 C24:0 0.5±0.4 0.6±0.1 0.3±0.1 0.4±0.1 0.4±0.01 0.4±0.1 0.6±0.3 0.2±0.1 1.1±0.4 n. d. 2.9±0.3 0.5±0.2 C16:1(n-7) 2.6±0.3 3.0±0.2 5.4±1.2 7.6±1.6 1.8±0.2 4.4±0.2 4.7±0.3 1.6±0.6 1.4±0.4 1.4±0.3 2.5±0.2 3.2±0.8 C16:1(n-9) n. d. n. d. n. d. n. d. 2.3±0.1 n. d. 1.3±0.2 2.6±1.1 1.4±0.2 n. d. n. d. n. d.C17:1(n-7) 0.3±0.1 n. d. 0.2±0.1 0.4±0.1 n. d. 0.4±0.3 n. d. 0.1±0.01 n. d. n. d. n. d. 0.5±0.2 C18:1(n-9) 6.8±1.0 7.4±0.3 24.2±9.8 7.6±1.9 9.4±0.8 6.6±1.4 6.2±0.4 7.6±0.5 6.6±1.2 1.4±0.1 3.9±0.2 4.4±0.6 C18:1(n-9) trans
n. d. n. d. 1.3±0.3 n. d. n. d. n. d. n. d. n. d. 1.1±0.1 1.4±0.2 4.3±0.1 2.7±0.1
C20:1(n-9) 1.2±0.3 1.9±0.1 1.1±0.1 1.8±0.4 0.2±0.01 1.7±0.7 n. d. n. d. n. d. n. d. n. d. n. d.C22:1(n-9 0.4±0.3 0.8±0.1 0.7±0.1 1.2±0. 2.2±0.5 0.6±0.1 n. d. 0.2±0.1 n. d. n. d. n. d. n. d.C18:2(n-6) 5.1±1.0 5.2±0.4 0.2±0.01 7.7±1.5 15.4±0.3 4.8±1.0 11±0.6 2.0±0.9 5.4±1.1 2.2±0.5 1.8±0.8 9.2±0.9 C18:3(n-6) 0.8±0.1 1.0±0.1 0.4±0.01 0.7±0.1 0.9±0.1 0.6±0.3 2.3±0.2 1.7±0.1 1.4±0.3 n. d. n. d. 0.6±0.3 C18:3(n-3) n. d. n. d. n. d. n. d. n. d. 2.9±2.0 n. d. n. d. n. d. n. d. n. d. n. d.C18:4(n-3) 11.5±2.8 12.3±0.9 0.9±1.1 8.8±1.8 4.3±0.3 6.9±3.9 11.6±1.4 23.5±0.4 6.5±1.6 n. d. n. d. n. d.C20:3(n6) 0.9±0.3 0.9±0.1 4.3±0.1 0.9±0.2 4.6±0.3 0.8±0.2 0.9±0.2 0.5±0.01 1.2±0.2 1.9±0.3 2.3±0.1 1.2±0.3 C20:3(n3) 0.9±0.4 0.7±0.1 0.6±0.1 1.3±0.2 0.5±0.01 1.0±0.2 1.2±0.4 1.3±0.2 n. d. n. d. n. d. n. d.C20:4(n-6) 14.1±4.5 18.0±0.3 2.3±1.2 20.9±5.1 16.9±0.9 17.6±4.8 15.4±0.1 13.6±0.6 9.4±1.3 58.3±0.5 7.3±0.1 17.6±4.2 C20:5(n-3) 3.1±0.7 6.4±0.7 14.9±1.0 4.7±1.4 0.4±0.3 4.2±1.6 5.2±0.4 9.3±0.1 5.9±1.5 0.3±0.1 n. d. n. d.C22:6(n-3) n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d.n.d.- not detected.
Table continued..
Appendix
250
FAs 61 62 63 64 65 66 67 68 69 70 71 C12:0 0.2±0.01 0.5±0.2 0.4±0.2 n. d. 0.4±0.01 n. d. 0.3±0.1 0.4±0.1 n. d. n. d. 0.9±0.3 C14:0 1.1±0.1 1.4±0.2 2.4±0.2 5.2±2.1 2.1±1.6 4.1±0.2 3.2±1.3 9.8±0.7 10±0.1 3.4±0. 8.6±1.6 C15:0 0.6±0.01 0.6±0.1 0.6±0.1 0.3±0.6 0.5±0.1 1.4±0.3 0.3±0.1 0.9±0.2 1.5±0.2 1.0±0.1 1.3±0.2 C16:0 25.1±2.3 49.5±3.8 54.9±1.9 31.5±3.3 39.6±2.9 46.8±3.4 21.6±2.7 44.5±4.0 50.3±1.5 53.1±2.6 38.1±3.4 C17:0 0.2±0.01 0.4±0.1 0.4±0.1 0.0 0.2±0.1 6.6±1.1 0.2±0.01 0.2±0.1 0.0 1.9±3.2 0.2±0.1 C18:0 2.3±0.2 4.8±2.0 2.3±1.7 3.1±1.0 2.2±0.4 3.9±0.8 12±8.1 2.1±0.9 2.1±0.2 4.0±2.7 10.9±0.2 C20:0 0.2±0.01 0.7±0.4 0.4±0.3 0.0 0.5±0.3 0.8±0.3 0.9±0.1 0.5±0.2 0.5±0.1 0.8±0.2 0.5±0.1 C22:0 0.4±0.01 1.3±0.7 0.4±0.2 0.0 0.5±0.3 0.5±0.1 0.7±0.4 0.5±0.3 0.7±0.2 1.1±0.5 0.6±0.1 C24:0 0.8±0.1 1.6±0.7 0.7±0.4 0.0 0.6±0.4 0.6±0.1 0.7±0.01 0.0 0.0 0.5±0.4 0.3±0.1 C16:1(n-7) 0.7±0.01 1.1±0.4 0.7±0.3 2.3±1.8 1.0±1.0 7.0±2.9 9.3±1.7 7.9±0.5 2.1±0.8 1.6±1.3 0.9±0.3 C16:1(n-9) n. d. n. d. n. d. 2.0±0.1 1.6±0.9 5.1±1.1 n. d. n. d. n. d. n. d. 4.4±1.0 C17:1(n-7) n. d. 0.3±0.1 0.3±0.1 n. d. 0.2±0.1 0.7±0.2 n. d. 0.1±0.1 n. d. n. d. n. d. C18:1(n-9) 1.1±0.01 2.3±1.5 0.4±0.1 2.5±1.3 2.6±0.7 3.0±0.6 3.1±2.7 3.6±0.2 5.0±0.4 1.0±0.4 6.1±0.1 C18:1(n-9) trans
1.4±0.1 4.8±0.6 3.4±0.6 4.9±3.0 3.0±0.8 2.0±0.7 1.8±0.2 6.6±0.6 7.9±1.5 3.6±0.7 4.3±0.3
C20:1(n-9) 0.3±0.01 0.6±0.3 0.2±0.1 n. d. 0.5±0.3 n. d. n. d. n. d. n. d. n. d. n. d. C22:1(n-9 0.0 0.1±0.1 0.2±0.1 n. d. 0.2±0.1 n. d. 5.4±0.4 n. d. n. d. n. d. 1.6±0.3 C18:2(n-6) 1.9±0.2 2.1±0.9 1.5±0.8 5.1±1.5 1.6±0.6 n. d. 23.2±4.2 2.2±0.3 2.3±0.8 11.4±2.4 n. d.C18:3(n-6) n. d. 0.8±0.4 n. d. n. d. 0.4±0.2 1.0±0.5 n. d. 0.3±0.1 n. d. n. d. n. d.C18:3(n-3) n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d.C18:4(n-3) n. d. n. d. n. d. n. d. 2.4±0.1 n. d. 11.8±1.8 n. d. n. d. n. d. n. d.C20:3(n6) 4.9±0.1 5.4±0.5 3.6±0.6 1.3±1.1 1.9±0.6 0.9±0.4 0.4±0.1 0.7±0.2 2.7±0.9 n. d. 0.6±0.3 C20:3(n3) n. d. 0.2±0.1 2.9±2.4 n. d. n. d. n. d. n. d. n. d. 2.8±0.8 n. d. n. d. C20:4(n-6) 58.8±2.5 20.8±4.0 23.9±1.6 41.1±3.1 12.9±2.5 10.7±3.8 5.0±0.8 7.3±2.6 5.6±0.8 15.1±1.2 8.7±1.0 C20:5(n-3) 0.0 0.5±0.2 0.4±0.3 0.8±0.3 25±3.7 5.0±3.0 n. d. 12.7±2.7 6.7±2.8 1.7±0.4 12.0±1.3 C22:6(n-3) n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d.n.d.- not detected.
Table continued..
Appendix
251
FAs 72 73 74 75 76 77 78 79 80 81 82 C12:0 0.3±0.1 0.3±0.1 0.9±0.1 0.3±0.1 0.5±0.3 0.5±0.3 n. d. 1.0±0.2 0.4±0.2 3.1±0.8 2.0±0.1 C14:0 0.9±0.1 1.0±0.1 1.8±0.1 1.2±0.3 1.1±0.2 4.5±0.7 4.6±0.8 7.0±0.7 6.4±1.3 9.3±4.3 11.3±0.7 C15:0 0.4±0.1 0.7±.1 0.7±0.1 0.9±0.1 0.8±0.1 0.5±0.3 1.7±0.2 0.9±0.3 0.8±0.1 2.7±0.9 0.9±0.1 C16:0 30.1±1.1 31±3.9 26.5±1.1 34.6±3.0 29.9±0.7 39.6±3.0 34.3±5.4 41.2±1.7 34.8±0.9 31±2.8 39.7±0.8 C17:0 0.2±0.01 0.5±0.1 0.4±0.1 0.4±0.2 0.6±0.3 0.4±0.2 1.0±0.1 0.4±0.1 0.3±1.2 1.1±0.8 0.4±0.1 C18:0 1.5±0.2 3.1±0.4 5.3±0.1 3.2±0.8 3.5±0.3 2.8±0.4 6.6±0.5 4.6±1.0 3.5±0.1 6.8±0.5 4.5±0.6 C20:0 0.2±0.01 0.2±0.2 0.2±0.1 0.2±0.1 0.4±0.2 0.2±0.01 0.7±0.1 0.3±0.01 0.2±0.3 0.3±0.3 0.2±0.01 C22:0 0.2±0.02 0.5±0.3 0.2±0.1 0.5±0.4 0.7±0.5 0.4±0.1 0.8±.1 0.5±0.2 0.8±0.2 1.8±0.3 0.5±0.1 C24:0 0.6±0.1 n. d. n. d. n. d. 0.00.8 0.4±.1 0.7±.1 0.7±0.3 0.4±0.7 0.8±0.3 0.3±0.1 C16:1(n-7) 0.7±0.1 0.3±0.1 0.5±0.1 1.3±0.9 1.1±0.1 1.5±.1 1.0±.1 3.1±0.6 4.1±0.1 3.6±3.0 3.0±0.2 C16:1(n-9) 0.7±0.1 0.8±0.1 1.6±0.3 0.6±0.1 0.8±0.1 n. d. 7.9±0.8 n. d. 3.2±0.3 n. d. 4.3±0.1 C17:1(n-7) 0.1±0.01 0.1±0.1 0.2±0.1 0.3±0.3 0.3±0.1 0.2±0.1 4.7±.1 n. d. 0.7±0.1 n. d. 0.2±0.1 C18:1(n-9) 1.2±0.1 2.9±0.4 5.7±1.0 1.8±1.3 2.3±0.8 2.4±0.2 1.1±0.2 4.5±0.8 4.4±1.0 3.8±1.8 3.5±0.4 C18:1(n-9) trans
1.0±0.1 3.2±0.4 2.1±0.1 2.2±1.5 2.1±0.1 3.7±0.2 1.8±0.1 6.3±0.7 13±1.3 2.5±0.3 4.3±0.3
C20:1(n-9) 0.7±0.2 1.4±0.3 1.6±0.1 1.7±0.2 1.4±0.1 n. d. n. d. n. d. n. d. n. d. 0.4±0.2 C22:1(n-9 0.8±0.1 1.4±0.3 1.3±0.2 1.3±0.3 1.4±0.5 n. d. n. d. 2.0±1.2 0.8±0.2 n. d. 1.3±0.6 C18:2(n-6) 2.7±1.2 4.9±0.5 4.3±0.5 3.7±0.1 4.3±1.0 4.5±1.3 1.3±0.2 7.9±1.7 6.1±1.6 24.8±0.8 6.7±0.5 C18:3(n-6) 0.3±0.1 0.7±0.3 0.7±0.1 0.8±0.2 1.6±0.3 0.6±0.1 n. d. n. d. 0.6±0.2 1.7±0.7 0.7±0.1 C18:3(n-3) n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d.C18:4(n-3) n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. 3.2±0.4 1.4±0.2 C20:3(n6) 2.4±0.6 2.6±0.2 3.7±0.2 2.7±0.2 3.7±0.4 2.3±0.1 2.2±1.0 0.5±0.1 0.6±0.2 0.0 0.4±0.1 C20:3(n3) 0.2±0.1 0.3±0.2 0.1±0.01 0.5±0.01 0.5±0.1 n. d. n. d. n. d. n. d. n. d. n. d.C20:4(n-6) 16.7±1.2 21±0.5 17.4±1.1 19.3±3.0 19.5±1.6 35±3.2 25.1±3.8 9.0±1.8 12.9±0.9 1.3±0.2 3.8±1.0 C20:5(n-3) 35.9±2.1 23.6±3.1 23.7±1.2 20.8±5.8 21.3±2.3 0.5±0.1 5.0±0.8 10.3±1.5 5.6±0.9 2.2±1.3 10±0.5 C22:6(n-3) n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d.n.d.- not detected.
Table continued..
Appendix
252
FAs 83 84 85 86 87 88 89 90 91 92 93 C12:0 0.3±0.2 0.5±0.1 0.3±0.2 1.7±0.1 0.8±0.2 0.5±0.3 0.5±0.1 1.0±0.6 0.3±0.1 3.0±0.9 3.1±0.8 C14:0 2.8±0.7 2.7±0.5 3.2±0.4 3.1±0.5 1.9±0.4 2.2±1.0 8.6±0.1 5.8±0.5 2.5±2.3 6.8±5.1 13.4±3.0 C15:0 0.7±0.1 0.5±0.3 0.8±0.1 1.1±0.4 0.8±0.5 0.5±0.2 0.5±0.1 0.9±0.1 0.3±0.01 0.7±0.1 0.7±0.2 C16:0 37.7±1.3 37±1.4 26.4±2.0 29.6±0.04 41.2±0.8 40±2.8 34.3±1.2 38.8±3.2 45.1±4.4 37±2.9 38.5±2.7 C17:0 0.2±0.1 0.4±0.3 0.2±0.1 0.1±0.01 0.7±0.4 0.2±0.1 0.2±0.1 0.3±0.1 n. d. n. d. 0.5±0.2 C18:0 3.3±0.8 2.1±0.7 1.7±0.2 3.5±0.1 3.2±0.5 2.6±0.4 1.9±0.3 3.5±0.5 1.0±0.1 2.6±0.8 3.0±0.9 C20:0 0.4±0.3 0.3±0.02 0.1±0.1 n. d. 0.4±0.1 0.4±0.2 0.2±0.1 0.2±0.1 0.2±0.1 n. d. 0.4±0.2 C22:0 0.6±0.1 0.6±0.1 0.2±0.1 0.2±0.1 0.5±0.1 0.3±0.01 0.2±0.1 0.5±0.3 0.1±0.1 n. d. 0.5±0.2 C24:0 1.5±1.7 0.6±0.4 0.1±0.01 0.0 0.4±0.3 0.5±0.4 0.0 0.2±0.1 0.2±0.1 n. d. 0.5±0.1 C16:1(n-7) 4.9±0.2 5.4±0.5 4.6±0.7 0.6±0.1 1.2±0.4 0.9±0.1 6.1±3.4 2.5±0.7 0.7±0.3 1.7±0.6 1.7±0.3 C16:1(n-9) n. d. n. d. n. d. n. d. n. d. 2.0±0.2 n. d. 1.5±0.9 n. d. 1.5±0.15 3.1±1.1 C17:1(n-7) n. d. 0.0 0.4±0.2 n. d. n. d. n. d. n. d. 0.6±0.1 n. d. n. d. n. d.C18:1(n-9) 2.4±0.1 2.1±0.2 3.1±0.5 3.4±0.5 3.5±0.3 2.6±0.7 2.6±0.8 3.5±0.4 2.9±1.2 3.3±0.6 3.4±1.0 C18:1(n-9) trans
6.0±1.8 4.2±1.2 16.9±2.1 4.1±1.7 8.3±1.0 4.8±1.0 4.5±1.4 3.7±0.4 0.9±0.2 4.9±1.0 3.5±1.0
C20:1(n-9) n. d. n. d. 0.3±0.2 n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d.C22:1(n-9 n. d. n. d. n. d. n. d. n. d. n. d. 0.4±0.3 n. d. n. d. n. d. n. d.C18:2(n-6) 1.9±1.5 2.8±0.3 2.2±0.2 5.8±1.0 3.8±0.5 3.9±1.0 2.9±0.2 3.7±0.4 0.3±0.1 3.1±0.4 n. d.C18:3(n-6) 1.1±0.6 1.1±0.3 1.4±1 1.3±0.8 0.0 1.6±0.9 0.7±0.3 0.4±0.2 n. d. n. d. n. d.C18:3(n-3) n. d. n. d. 0.9±0.3 n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d.C18:4(n-3) 1.6±.07 0.8±0.4 0.6±0.6 n. d. n. d. 2.1± 1.0±0.1 0.8±0.1 n. d. n. d. n. d.C20:3(n6) 3.7±0.5 1.4±1.0 0.8±0.2 1.0±0.3 0.7±0.2 0.5±0.3 0.3±0.1 1.1±0.2 0.3±0.1 3.6±2.3 1.0±0.5 C20:3(n3) n. d. n. d. 0.2±0.1 n. d. n. d. n. d. n. d. 0.3±0.1 n. d. n. d. n. d.C20:4(n-6) 10.8±5.4 11.1±2.7 35.4±1.9 25.2±6.3 19.8±0.5 8.2±2.2 15.5±1.1 9.2±1.5 20.6±4.4 18.2±2.2 14.3±2.1 C20:5(n-3) 20.2±6.8 26.4±3.6 0.4±0.3 19.4±4.1 12.9±1.1 25.8±6.1 19.6±4.3 21.2±2.7 24.8±1.5 14.1±1.4 12.5±1.9 C22:6(n-3) n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d. n. d.n.d.- not detected.
Table continued..
Appendix
253
FAs 94 95 96 97 98 99 100 LSD (0.1%)A C12:0 0.5±0.5 1.2±0.3 0.6±0.1 1.0±0.1 0.8±0.4 0.7±0.8 2.3±0.8 0.9 C14:0 8.0±2.5 7.5±5.6 11.9±0.9 4.2±0.4 4.3±0.8 5.0±0.7 5.3±0.6 4.2 C15:0 0.4±0.3 1.0±0.1 1.3±0.1 1.0±0.4 1.5±1.0 1.3±0.2 1.4±0.4 1.2 C16:0 34.3±1.8 45.8±15.4 56.6±2.7 37.5±1.1 36.9±2.9 54.3±3.7 44.2±4.8 10.7 C17:0 0.2±0.01 0.3±0.1 0.3±0.01 0.6±0.1 0.9±0.6 0.4±0.1 2.6±2.6 1.1 C18:0 3.9±2.9 4.5±1.6 2.7±0.8 3.6±1.3 4.3±0.4 3.3±1.8 7.5±2.1 4.4 C20:0 0.2±0.01 0.4±0.2 0.3±0.02 0.6±0.3 1.2±0.3 0.2±0.1 0.4±0.1 0.9 C22:0 0.3±0.1 0.7±0.4 0.5±0.1 1.2±0.6 1.6±0.4 0.3±0.01 1.0±0.01 0.8 C24:0 0.5±0.2 0.6±0.1 0.3±0.1 1.8±0.1 1.1±0.4 0.3±0.01 0.8±0.2 1.5 C16:1(n-7) 4.7±0.8 2.5±1.3 3.3±0.4 1.8±0.5 2.5±0.3 2.0±0.1 1.1±0.1 4.6 C16:1(n-9) 1.7±0.7 3.5±1.3 1.7±1.7 3.3±0.2 4.5±1.8 2.9±0.4 6.7±2.4 3.9 C17:1(n-7) n. d. 0.1±0.01 0.5±0.2 n. d. n. d. n. d. 0.7±0.1 0.4 C18:1(n-9) 3.7±0.9 3.8±1.2 4.1±0.5 2.8±0.3 3.4±1.4 2.8±1.0 4.0±0.4 4.0 C18:1(n-9) trans
4.4±0.8 3.7±0.8 4.9±0.3 6.3±1.6 7.2±1.7 3.4±1.1 3.9±1.9 2.1
C20:1(n-9) n. d. n. d. n. d. n. d. 2.0±0.5 0.6±0.1 0.9±0.1 0.5 C22:1(n-9 0.1±0.01 n. d. 0.4±0.1 n. d. n. d. n. d. 0.9±0.1 0.7 C18:2(n-6) 4.4±0.7 2.4±0.7 1.8±0.1 6.7±1.1 8.1±0.7 6.1±2.1 4.9±1.2 3.8 C18:3(n-6) 0.1±0.01 0.0 0.3±0.1 1.3±0.4 0.8±0.6 n. d. 1.3±0.3 2.3 C18:3(n-3) 0.0 n. d. n. d. n. d. n. d. n. d. n. d. 6.4 C18:4(n-3) 1.4±0.1 n. d. n. d. n. d. 1.9±0.8 n. d. 1.8±0.2 5.6 C20:3(n6) 0.2±0.1 0.5±0.3 0.2±0.1 1.4±0.4 1.4±0.4 0.5±0.1 0.5±0.1 2.0 C20:3(n3) n. d. n. d. n. d. n. d. n. d. n. d. 0.3±0.1 1.5 C20:4(n-6) 14.6±0.9 12.1±5.0 4.2±0.3 2.8±0.6 4.4±0.9 9.2±3.1 1.2±0.8 5.3 C20:5(n-3) 16.2±2.2 9.4±2.0 3.9±0.1 22.2±1.5 10.9±2.1 5.4±1.3 5.1±1.7 5.2 C22:6(n-3) n. d. n. d. n. d. n. d. n. d. n. d. n. d. 1.2 n.d.- not detected; A: LSD values obtained from ANOVA, The values in a row are significantly different at p≤0.001.
Appendix
254
Appendix IV
Percent relative abundance of polar lipid molecular species present in different macroalgae as determined by ESI-MS scans.
Mass Compound name UL UF UT UB UTb GC GCY GFO GD GS GTx GFU PT ST CI SA
766.1 MGDG (34:5) 28 98 58 28 98 10 8 10 8 8 18 5 10 8 32 12 768.1 MGDG (34:4) 70 50 45 22 28 15 10 12 10 5 8 8 12 10 24 15 770.5 MGDG (34:3) 27 8 20 20 18 20 8 10 18 5 12 5 15 6 22 24 772 MGDG (34:2) 20 15 22 10 8 10 5 8 15 5 8 5 28 15 18 20 794 MGDG (36:8) 8 5 8 21 5 15 10 8 12 5 18 5 95 10 15 18 796 MGDG (36:6) 6 5 5 12 5 12 8 8 10 5 8 5 98 80 12 24 823 MGDG (38:5) 5 5 8 14 5 18 10 15 8 5 12 12 5 6 16 98 843.7 MGDG (40:9) 8 6 5 98 5 72 88 9 88 30 5 5 8 15 12 12 845 MGDG (40:8) 5 5 15 15 5 22 12 80 10 8 18 5 5 10 18 6 908.6 DGDG (32:3) 8 5 5 10 5 8 10 30 12 8 12 5 5 8 5 5 910 DGDG (32:2) 5 10 15 22 5 70 60 48 10 8 15 5 8 12 15 24 931 DGDG (34:6) 10 5 15 28 5 18 15 12 12 15 12 5 8 50 32 5 941.3 DGDG (34:1) 22 12 25 28 5 48 68 60 22 12 24 8 8 10 12 6 958 DGDG (36:8) 12 5 5 22 5 10 15 18 15 5 22 5 8 6 22 6 960 DGDG (36:6) 18 5 10 22 5 8 10 12 18 5 8 5 5 10 18 6 962 DGDG (36:5) 11 5 8 18 5 18 12 15 18 5 5 5 15 28 15 6 968 DGDG (36:4) 68 5 32 60 5 98 50 45 68 78 8 5 10 10 12 8 1004.6 DGDG (40:9) 5 5 8 10 5 12 12 12 12 8 70 5 5 6 8 5 1006.6 DGDG (40:8) 5 6 8 10 5 52 55 52 52 18 10 8 6 8 10 6 761.34 SQDG (30:2) 22 18 12 5 5 20 5 10 30 10 8 12 5 12 18 12 765.77 SQDG (30:0) 18 12 11 5 5 18 5 8 32 8 5 8 40 51 40 38 785.86 SQDG (32:4) 30 70 15 10 30 15 5 5 18 8 8 5 30 10 20 32 791.5 SQDG (32:1) 15 10 10 5 8 22 5 8 52 10 8 18 80 98 85 98 793.84 SQDG (32:0) 12 12 60 5 8 12 5 8 50 18 22 60 35 78 80 56 815.37 SQDG (34:3) 40 98 35 12 28 25 5 5 22 8 10 40 15 30 20 28 819 SQDG (38:9) 12 92 98 10 8 22 5 5 18 5 18 15 28 98 97 85 821 SQDG (38:8) 45 30 35 10 5 18 5 5 30 5 8 10 15 48 25 42 704.5 DGTS (32:4) 6 4 6 8 5 0 0 0 0 0 0 0 0 0 0 0 706.5 DGTS (32:3) 8 6 8 8 5 0 0 0 0 0 0 0 0 0 0 0 710.5 DGTS (32:1) 10 8 8 18 5 0 0 0 0 0 0 0 0 0 0 0 732 DGTS (34:4) 6 10 10 15 5 0 0 0 0 0 0 0 0 0 0 0 734 DGTS (34:3) 6 12 11 18 5 0 0 0 0 0 0 0 0 0 0 0 736.5 DGTS (34:2) 8 12 12 22 5 0 0 0 0 0 0 0 0 0 0 0 738.5 DGTS (34:1) 18 20 18 60 5 0 0 0 0 0 0 0 0 0 0 0 762 DGTS (36:3) 20 18 15 95 10 0 0 0 0 0 0 0 0 0 0 0 764 DGTS (36:2) 30 25 20 25 20 0 0 0 0 0 0 0 0 0 0 0 676.49 PC(28:1) 0 0 0 0 10 18 28 40 36 8 5 5 0 0 0 0 678.5 PC(28:0) 0 0 0 0 5 8 16 12 4 4 12 18 0 0 0 0 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
255
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
724.49 PC(32:5) 0 0 0 0 0 2 12 14 16 5 2 2 0 0 0 0
726.5 PC(32:4) 0 0 0 0 0 2 9 12 8 2 5 5 0 0 0 0
728.52 PC(32:3) 0 0 0 0 0 2 2 2 5 2 4 5 0 0 0 0
730.53 PC(32:2) 0 0 0 0 0 2 2 2 4 2 4 5 0 0 0 0
732.55 PC(32:1) 0 0 0 0 35 2 2 2 4 2 4 5 0 0 0 0
734.56 PC(32:0) 0 0 0 0 98 2 2 2 4 2 4 5 0 0 0 0
746.56 PC(33:1) 0 0 0 0 4 0 0 0 0 0 0 0 0 0 0 0
748.58 PC(33:0) 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0
750.5 PC(34:6) 0 0 0 0 2 2 2 2 2 2 2 2 0 0 0 0
752.25 PC(34:5) 0 0 0 0 2 2 2 2 2 2 2 3 0 0 0 0
754.53 PC(34:4) 0 0 0 0 20 8 6 2 2 2 2 5 0 0 0 0
756.55 PC(34:3) 0 0 0 0 80 6 6 2 2 2 2 4 0 0 0 0
758.56 PC(34:2) 0 0 0 0 48 4 6 2 2 2 2 4 0 0 0 0
760.58 PC(34:1) 0 0 0 0 22 2 6 2 2 2 2 4 0 0 0 0
762.6 PC(34:0) 0 0 0 0 12 2 4 2 2 2 2 5 0 0 0 0
774.5 PC(36:8) 0 0 0 0 4 2 10 2 2 2 2 2 0 0 0 0
776.5 PC(36:7) 0 0 0 0 4 0 0 0 0 0 0 0 0 0 0 0
778.5 PC(36:6) 0 0 0 0 4 0 0 0 0 0 0 0 0 0 0 0
780.5 PC(36:5) 0 0 0 0 4 2 4 80 20 8 2 2 0 0 0 0
782.56 PC(36:4) 0 0 0 0 12 2 4 6 4 4 3 5 0 0 0 0
784.5 PC(36:3) 0 0 0 0 10 2 4 4 4 2 3 4 0 0 0 0
786.6 PC(36:2) 0 0 0 0 8 2 4 5 4 2 3 4 0 0 0 0
788.6 PC(36:1) 0 0 0 0 6 3 4 4 4 2 3 4 0 0 0 0
790.6 PC(36:0) 0 0 0 0 12 6 4 6 4 2 3 4 0 0 0 0
800.52 PC(38:9) 0 0 0 0 8 5 4 6 18 17 3 4 0 0 0 0
802.5 PC(38:8) 0 0 0 0 0 8 98 36 12 8 4 3 0 0 0 0
804.55 PC(38:7) 0 0 0 0 0 10 27 24 4 6 2 2 0 0 0 0
806.56 PC(38:6) 0 0 0 0 0 5 16 12 2 5 2 2 0 0 0 0 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
256
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
808.5 PC(38:5) 0 0 0 0 0 5 8 5 2 5 2 2 0 0 0 0
810.6 PC(38:4) 0 0 0 0 10 0 0 0 0 0 0 0 0 0 0 0
812.6 PC(38:3) 0 0 0 0 6 0 0 0 0 0 0 0 0 0 0 0
814.6 PC(38:2) 0 0 0 0 4 0 0 0 0 0 0 0 0 0 0 0
816.64 PC(38:1) 0 0 0 0 2 2 2 15 2 2 2 2 0 0 0 0
818.6 PC(38:0) 0 0 0 0 4 2 2 8 2 2 2 2 0 0 0 0
826.53 PC(40:10) 0 0 0 0 0 2 2 6 4 2 4 2 0 0 0 0
828.5 PC(40:9) 0 0 0 0 0 2 2 4 4 2 4 3 0 0 0 0
830.5 PC(40:8) 0 0 0 0 0 2 2 4 4 2 2 3 0 0 0 0
832.5 PC(40:7) 0 0 0 0 0 2 2 4 4 2 2 4 0 0 0 0
834.6 PC(40:6) 0 0 0 0 0 2 2 4 4 2 4 2 0 0 0 0
836.6 PC(40:5) 0 0 0 0 0 2 2 4 4 2 5 4 0 0 0 0
838.63 PC(40:4) 0 0 0 0 0 2 2 4 4 2 5 8 0 0 0 0
840.64 PC(40:3) 0 0 0 0 4 2 2 8 5 2 5 5 0 0 0 0
842.6 PC(40:2) 0 0 0 0 0 2 2 6 5 2 4 0 0 0 0 0
844.6 PC(40:1) 0 0 0 0 0 2 2 6 5 2 4 0 0 0 0 0
846.6 PC(40:0) 0 0 0 0 0 2 2 4 5 2 4 0 0 0 0 0
852.5 PC(42:11) 0 0 0 0 0 2 2 20 10 8 2 2 0 0 0 0
854.56 PC(42:10) 0 0 0 0 0 2 2 8 6 6 2 2 0 0 0 0
856.5 PC(42:9) 0 0 0 0 0 2 2 6 5 4 2 2 0 0 0 0
858.6 PC(42:8) 0 0 0 0 0 2 2 4 4 3 2 2 0 0 0 0
860.6 PC(42:7) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
862.63 PC(42:6) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
864.6 PC(42:5) 0 0 0 0 4 2 2 25 2 3 2 2 0 0 0 0
866.6 PC(42:4) 0 0 0 0 4 2 2 12 2 3 2 2 0 0 0 0
874.72 PC(42:0) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
892.67 PC(44:5) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
440.0 LPC(12:0) 0 0 0 0 0 8 12 20 12 6 12 15 0 0 0 0 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
257
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
468.0 LPE(14:0) 0 0 0 0 0 2 8 8 5 2 6 5 0 0 0 0
494.32 LPC(16:1) 0 0 0 0 12 0 0 0 0 0 0 0 0 0 0 0
496.34 LPC(16:0) 0 0 0 0 8 0 0 0 0 0 0 0 0 0 0 0
508.33 LPC(17:1) 0 0 0 0 6 0 0 0 0 0 0 0 0 0 0 0
542.32 LPC(20:5) 0 0 0 0 0 2 2 12 15 2 2 2 0 0 0 0
544.33 LPC(20:4) 0 0 0 0 0 2 2 6 8 4 4 2 0 0 0 0
546.35 LPC(20:3) 0 0 0 0 0 2 2 5 6 5 4 2 0 0 0 0
580.39 PE(24:0) 4 4 5 4 0 2 2 2 4 2 5 5 4 4 5 4
660.45 PE(30:2) 0 0 0 0 0 0 0 0 0 0 0 0 5 5 6 6
676.49 PE(31:1) 6 5 6 98 12 17 26 2 4 8 12 6 7 5 6 4
678.5 PE(31:0) 5 4 4 28 7 12 22 2 5 4 5 15 13 12 15 11
682.44 PE(32:5) 4 2 2 4 2 6 8 2 5 4 4 4 5 4 4 3
684.45 PE(32:4) 4 2 2 2 2 2 6 2 5 4 28 4 5 4 4 3
686.47 PE(32:3) 0 0 0 0 0 2 5 2 4 4 15 4 12 12 14 10
688.49 PE(32:2) 0 0 0 0 0 2 5 8 21 4 8 2 11 12 10 8
690.5 PE(32:1) 0 0 0 0 0 2 5 5 10 4 4 2 5 6 4 4
692.52 PE(32:0) 0 0 0 0 0 2 5 5 8 3 4 2 15 15 14 12
704.5 PE(33:1) 0 0 0 0 0 0 0 0 0 0 0 0 4 5 4 2
706.53 PE(33:0) 0 0 0 0 0 0 0 0 0 0 0 0 3 5 5 2
708.45 PE(34:6) 0 0 0 0 0 0 0 0 0 0 0 0 3 4 5 3
710.47 PE(34:5) 4 4 2 2 10 0 0 0 0 0 0 0 3 4 5 4
712.49 PE(34:4) 4 4 2 2 4 2 4 4 4 3 4 4 3 6 5 2
714.5 PE(34:3) 4 4 2 2 4 2 4 10 5 3 4 4 6 7 8 6
716.52 PE(34:2) 0 0 0 0 0 2 4 8 4 2 10 4 4 5 6 4
718.54 PE(34:1) 0 0 0 0 0 2 4 6 4 2 4 5 4 5 6 5
720.55 PE(34:0) 0 0 0 0 0 2 6 4 2 2 2 2 4 5 6 5
732.45 PE(36:8) 5 5 6 2 30 2 6 2 2 2 2 2 4 5 7 6
734.47 PE(36:7) 5 4 7 2 98 2 5 2 8 2 4 4 4 5 6 6 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
258
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
736.49 PE(36:6) 5 4 7 2 55 2 5 2 7 2 4 4 4 5 6 4
738.5 PE(36:5) 5 3 6 2 20 2 5 2 4 2 6 5 4 5 7 4
740.52 PE(36:4) 4 3 5 2 10 2 5 2 4 2 5 4 4 5 4 4
742.5 PE(36:3) 2 2 2 3 4 2 5 2 5 2 6 4 2 5 4 4
744.45 PE(36:2) 2 2 2 10 5 2 5 2 4 2 6 4 2 4 4 3
746.56 PE(36:1) 2 2 2 8 2 0 0 0 0 0 0 0 2 4 2 2
748.58 PE(36:0) 2 2 2 2 2 0 0 0 0 0 0 0 2 2 2 2
758.47 PE(38:9) 6 4 16 12 40 0 0 0 0 0 0 0 6 10 8 6
760.49 PE(38:8) 5 5 12 4 20 0 0 0 0 0 0 0 0 0 0 0
762.5 PE(38:7) 0 0 0 5 10 0 0 0 0 0 0 0 0 0 0 0
764.52 PE(38:6) 2 2 2 14 6 0 0 0 0 0 0 0 0 0 6 5
766.5 PE(38:5) 2 2 2 8 6 0 0 0 0 0 0 0 5 4 5 4
768.5 PE(38:4) 2 2 2 5 18 0 0 0 0 0 0 0 7 8 6 4
770.56 PE(38:3) 2 2 2 5 6 0 0 0 0 0 0 0 4 5 4 4
772.58 PE(38:2) 0 0 0 0 0 0 0 0 0 0 0 0 3 5 3 3
774.6 PE(38:1) 0 0 0 0 0 0 0 0 0 0 0 0 2 4 4 2
776.6 PE(38:0) 2 2 4 4 5 2 8 2 4 2 3 3 4 4 4 2
784.49 PE(40:10) 0 0 0 0 0 2 6 2 4 2 4 2 15 16 20 10
786.5 PE(40:9) 0 0 0 0 0 2 6 2 3 2 3 5 6 8 8 4
788.5 PE(40:8) 0 0 0 0 0 2 5 2 3 2 4 5 6 6 6 4
790.5 PE(40:7) 0 0 0 0 0 2 5 18 22 2 3 3 5 4 4 5
792.5 PE(40:6) 0 0 0 0 0 2 5 98 98 2 4 4 4 2 2 4
794.56 PE(40:5) 0 0 0 0 0 2 5 35 36 2 2 2 2 2 2 4
796.58 PE(40:4) 0 0 0 0 0 2 5 12 24 2 2 2 2 2 2 3
798.6 PE(40:3) 0 0 0 0 0 2 5 8 6 2 2 2 2 2 2 3
800.6 PE(40:2) 0 0 0 0 0 8 5 4 2 14 4 4 2 2 2 3
802.6 PE(40:1) 0 0 0 0 0 4 90 4 3 8 3 3 2 2 4 3
804.6 PE(40:0) 4 4 4 4 4 10 30 3 3 4 4 5 2 2 4 3 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
259
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
810.5 PE(42:11) 0 0 0 0 0 6 12 3 3 3 3 3 2 2 4 3
812.5 PE(42:10) 0 0 0 0 0 5 12 10 27 2 2 2 2 2 4 2
814.5 PE(42:9) 0 0 0 0 0 4 10 6 24 2 2 2 2 2 4 2
816.5 PE(42:8) 0 0 0 0 0 4 8 8 10 2 2 2 2 2 4 2
818.5 PE(42:7) 4 4 2 2 2 2 8 5 4 2 2 2 2 2 4 2
820.58 PE(42:6) 4 2 2 2 2 3 4 5 5 2 2 2 2 2 4 2
822.6 PE(42:5) 4 2 3 4 2 3 75 2 2 20 2 2 2 2 4 4
824.6 PE(42:4) 4 2 3 4 2 2 36 2 2 10 2 2 2 2 4 4
832.67 PE(42:0) 0 0 0 0 0 0 0 0 0 0 0 0 10 15 16 12
850.63 PE(44:5) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
852.6 PE(44:4) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
426.2 LPE(14:0) 0 0 0 0 0 0 0 0 0 0 0 0 8 8 12 8
440 LPE(15:0) 0 0 0 0 0 0 0 0 0 0 0 0 7 8 10 6
452 LPE(16:1) 0 0 0 0 0 0 0 0 0 0 0 0 4 6 5 3
454 LPE(16:0) 0 0 0 0 0 0 0 0 0 0 0 0 4 4 5 3
426.2 LPE(14:0) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0
440 LPE(15:0) 0 0 0 0 0 4 12 4 5 6 10 15 0 0 0 0
452 LPE(16:1) 0 0 0 0 0 4 9 2 22 2 6 10 0 0 0 0
454 LPE(16:0) 0 0 0 0 0 4 8 2 8 2 6 5 0 0 0 0
508.3 LPE(20:1) 6 5 22 12 20 0 0 0 0 0 0 0 0 0 0 0
609.37 PG(24:0) 4 8 4 12 2 2 2 2 4 2 4 4 4 2 2 2
635.39 PG(26:1) 2 4 2 4 2 0 0 0 0 0 0 0 0 0 0 0
665.4 PG(28:0) 0 0 0 0 0 2 2 2 4 2 4 4 2 2 2 2
689.43 PG(30:2) 8 6 5 14 4 4 5 8 16 4 5 10 4 2 2 4
691.4 PG(30:1) 6 5 3 48 15 0 4 4 10 5 22 5 4 2 2 5
693.4 PG(30:0) 6 2 2 14 8 2 2 2 6 4 12 5 5 2 2 3 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
260
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
705.47 PG(31:2) 5 2 2 2 2 0 0 0 0 0 0 0 4 2 2 2
707.47 PG(31:1) 5 2 2 2 2 0 0 0 0 0 0 0 5 2 2 2
709.4 PG(31:0) 0 0 0 0 0 0 0 0 0 0 0 0 4 2 2 2
711.42 PG(32:5) 0 0 0 0 0 0 0 0 0 0 0 0 6 2 2 2
713.4 PG(32:4) 0 0 0 0 0 0 0 0 0 0 0 0 5 2 2 2
715.4 PG(32:3) 0 0 0 0 0 0 0 0 0 0 0 0 4 2 2 2
717.4 PG(32:2) 0 0 0 0 0 0 0 0 0 0 0 0 4 2 2 2
719.4 PG(32:1) 0 0 0 0 0 2 4 5 5 2 50 6 4 2 2 2
721.4 PG(32:0) 0 0 0 0 0 2 2 2 5 2 30 5 4 2 2 2
733.5 PG(33:1) 4 4 2 2 2 2 2 2 5 3 15 4 5 2 2 4
735.5 PG(33:0) 4 4 2 2 2 2 2 2 5 4 12 5 5 2 2 2
737.4 PG(34:6) 4 12 2 2 6 2 2 2 6 3 10 4 5 2 4 3
739.4 PG(34:5) 8 10 5 4 6 2 2 2 5 3 8 4 5 2 3 4
741.4 PG(34:4) 7 8 20 5 25 2 2 2 4 2 4 3 5 4 4 4
743.4 PG(34:3) 8 6 16 4 18 2 2 2 4 2 5 3 5 5 2 4
745.4 PG(34:2) 6 6 6 3 15 2 2 2 4 2 16 3 5 3 2 4
747.4 PG(34:1) 5 7 5 3 8 2 2 2 4 2 11 0 5 2 2 5
749.4 PG(34:0) 4 8 4 3 5 0 2 2 4 4 9 0 4 4 4 4
761.4 PG(36:8) 5 2 2 2 2 0 0 0 0 0 0 0 4 5 6 6
763.4 PG(36:7) 5 2 2 2 2 0 0 0 0 0 0 0 10 6 15 7
765.4 PG(36:6) 5 6 2 8 2 18 14 20 20 16 14 25 33 18 45 25
767.4 PG(36:5) 5 7 22 4 12 8 8 8 6 8 8 12 18 19 16 11
769.4 PG(36:4) 5 8 2 4 2 7 6 6 5 6 7 8 14 8 8 8
771.4 PG(36:3) 5 15 2 4 6 6 5 4 5 4 4 4 8 4 6 2
773.4 PG(36:2) 5 12 2 4 6 5 4 4 4 4 4 4 0 0 0 0
775.4 PG(36:1) 5 10 2 4 6 0 0 0 0 0 0 0 0 0 0 0
791.4 PG(38:7) 5 8 2 3 6 2 2 2 23 11 8 4 40 25 50 44
793.4 PG(38:6) 78 98 98 48 42 98 98 98 90 75 6 98 27 30 74 36 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
261
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
795.4 PG(38:5) 25 30 35 15 12 32 41 30 40 36 98 26 16 15 33 24
797.4 PG(38:4) 12 16 80 12 8 17 12 12 20 24 50 10 11 12 16 12
799.4 PG(38:3) 6 7 5 8 5 12 8 8 10 17 28 5 7 4 8 8
801.4 PG(38:2) 4 4 2 4 4 8 6 4 2 2 8 4 5 4 4 6
803.4 PG(38:1) 4 4 2 2 2 6 4 2 2 2 6 3 4 4 5 4
805.5 PG(38:0) 0 0 0 0 0 4 4 2 2 2 5 4 4 4 4 6
813.47 PG(40:10) 24 6 12 12 12 4 39 40 17 10 10 4 22 12 15 6
815.47 PG(40:9) 16 12 14 10 98 8 21 18 15 8 20 5 16 15 16 11
817.5 PG(40:8) 12 18 10 6 33 4 22 12 14 6 11 4 12 16 18 14
819.5 PG(40:7) 98 97 80 18 30 4 10 8 6 5 10 4 28 52 98 98
821.5 PG(40:6) 34 34 25 6 16 4 8 6 6 4 8 5 16 22 30 38
823.5 PG(40:5) 12 14 8 5 8 2 5 4 4 3 7 5 14 14 11 16
825.5 PG(40:4) 8 10 8 18 8 2 5 4 4 3 6 5 8 8 8 8
827.5 PG(40:3) 3 6 6 12 9 2 5 4 4 3 5 4 7 6 5 7
829.5 PG(40:2) 3 5 4 6 2 2 5 4 4 3 4 0 5 4 4 4
831.5 PG(40:1) 4 5 4 4 2 2 5 4 2 3 4 0 5 8 20 5
833.5 PG(40:0) 4 5 4 4 2 2 5 4 2 3 4 0 5 6 16 5
839.48 PG(42:11) 4 4 2 4 2 12 5 4 2 4 4 0 4 8 8 5
841.48 PG(42:10) 4 4 2 2 2 10 36 42 30 31 14 12 4 7 6 6
843.48 PG(42:9) 4 4 2 2 2 8 15 26 15 22 10 10 4 6 7 4
845.48 PG(42:8) 4 4 2 2 2 5 8 12 8 16 9 8 4 5 8 3
847.48 PG(42:7) 4 4 2 2 2 4 9 8 6 10 6 6 0 0 0 0
849.48 PG(42:6) 4 4 2 2 2 4 6 8 6 4 6 6 0 0 0 0
851.48 PG(42:5) 4 4 2 5 2 2 6 6 4 3 5 5 0 0 0 0
853.48 PG(42:4) 4 4 2 8 2 2 4 6 4 2 5 5 0 0 0 0
861.65 PG(42:0) 0 0 0 0 0 2 28 34 25 2 4 4 0 0 0 0
889.6 PG(44:0) 0 0 0 0 0 2 2 2 12 5 13 10 0 0 0 0
455.24 LPG(14:0) 0 0 0 0 0 4 2 2 2 2 3 4 0 0 0 0 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
262
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
481.25 LPG(16:1) 4 8 4 2 4 0 0 0 0 0 0 0 0 0 0 0
483.27 LPG(16:0) 4 12 4 4 2 0 0 0 0 0 0 0 0 0 0 0
511.3 LPG(18:0) 0 0 0 0 0 0 0 0 0 0 0 0 8 7 6 5
529.25 LPG(20:5) 8 12 5 4 4 6 4 5 8 18 8 8 20 10 12 7
531.27 LPG(20:4) 0 0 0 0 0 4 4 6 7 7 10 4 0 0 0 0
622.37 PS(24:0) 2 4 2 2 4 2 2 2 4 2 4 4 4 3 2 2
648.38 PS(26:1) 0 0 0 0 0 0 0 0 0 0 0 0 4 2 3 2
650.4 PS(26:0) 0 0 0 0 0 4 2 2 4 2 4 5 4 2 5 4
674.4 PS(28:2) 0 0 0 0 0 4 3 2 4 2 4 4 6 2 2 4
676.41 PS(28:1) 0 0 0 0 0 0 0 0 0 0 0 0 8 2 2 2
678.4 PS(28:0) 0 0 0 0 0 2 5 2 4 2 4 4 4 2 2 2
690.4 PS(29:1) 0 0 0 0 0 2 5 6 17 4 18 8 5 3 2 2
692.4 PS(29:0) 0 0 0 0 0 2 4 4 10 5 12 5 5 3 2 2
702.43 PS(30:2) 4 6 2 8 4 2 2 2 5 2 4 5 4 3 2 3
704.4 PS(30:1) 0 0 0 0 0 2 2 2 6 2 4 4 4 3 2 3
706.4 PS(30:0) 0 0 0 0 0 2 2 2 5 2 4 5 6 2 2 3
716.45 PS(31:2) 0 0 0 0 0 2 8 5 4 5 10 4 6 3 2 3
718.45 PS(31:1) 0 0 0 0 0 2 6 3 5 4 51 8 5 2 2 2
720.48 PS(31:0) 0 0 0 0 0 2 8 2 4 2 22 5 5 4 2 3
724.4 PS(32:5) 0 0 0 0 0 2 2 2 5 2 27 4 4 4 4 3
726.4 PS(32:4) 0 0 0 0 0 2 2 2 6 2 5 4 4 4 4 4
728.4 PS(32:3) 0 0 0 0 0 2 2 2 4 2 5 4 4 4 4 4
730.4 PS(32:2) 0 0 0 0 0 2 4 2 4 6 75 22 4 2 3 2
732.4 PS(32:1) 0 0 0 0 0 2 4 2 4 4 38 11 5 2 3 2
734.4 PS(32:0) 0 0 0 0 0 2 4 2 4 2 20 10 4 2 3 2
746.5 PS(33:1) 7 4 5 4 5 2 3 2 4 2 17 8 4 2 3 4
748.5 PS(33:0) 8 5 2 2 4 2 3 2 4 2 14 6 4 2 3 5
750.4 PS(34:6) 8 5 2 2 2 2 3 2 4 2 12 2 5 2 2 3 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
263
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
752.4 PS(34:5) 4 5 2 4 2 2 3 2 4 2 10 3 5 2 3 3
754.4 PS(34:4) 5 5 2 4 2 2 3 2 4 2 5 2 5 2 3 2
756.4 PS(34:3) 4 5 2 2 2 2 4 2 4 2 4 2 5 2 3 2
758.4 PS(34:2) 4 5 2 2 2 2 4 2 4 2 4 2 5 2 2 2
760.4 PS(34:1) 7 4 2 2 2 2 4 2 4 2 4 2 5 2 5 2
762.4 PS(34:0) 8 4 2 2 2 2 4 2 4 2 4 2 5 2 4 8
774.4 PS(36:8) 4 12 2 2 4 0 0 0 0 0 0 0 8 2 4 2
776.4 PS(36:7) 4 11 2 2 5 0 0 0 0 0 0 0 8 2 5 2
778.4 PS(36:6) 4 8 4 2 16 0 0 0 0 0 0 0 8 2 4 2
780.4 PS(36:5) 4 6 5 2 8 0 0 0 0 0 0 0 8 2 5 6
782.4 PS(36:4) 2 4 5 2 6 0 0 0 0 0 0 0 8 4 8 7
784.4 PS(36:3) 2 4 2 2 4 0 0 0 0 0 0 0 7 4 10 6
786.4 PS(36:2) 2 4 2 2 4 0 0 0 0 0 0 0 7 2 7 6
788.4 PS(36:1) 2 4 2 2 4 0 0 0 0 0 0 0 8 4 8 6
790.5 PS(36:0) 2 4 2 4 2 5 2 8 22 2 4 2 5 5 50 45
800.45 PS(38:9) 2 4 3 6 8 4 2 2 2 2 4 2 4 6 12 33
802.4 PS(38:8) 2 4 2 4 4 4 2 2 2 2 5 2 4 2 8 5
804.4 PS(38:7) 2 4 2 5 5 3 2 2 2 2 5 2 4 2 6 6
806.4 PS(38:6) 2 5 2 2 2 3 2 2 2 2 4 2 4 2 5 8
808.4 PS(38:5) 2 5 2 4 2 3 2 2 2 2 6 2 16 2 5 6
810.4 PS(38:4) 8 4 4 4 2 2 2 2 2 2 6 4 12 2 5 8
812.4 PS(38:3) 10 4 4 4 8 2 12 40 16 12 6 5 10 2 4 9
814.4 PS(38:2) 12 12 12 4 98 8 10 15 10 8 15 5 14 2 4 11
816.4 PS(38:1) 24 18 56 4 30 5 8 18 15 7 7 5 8 2 4 12
818.5 PS(38:0) 25 12 72 16 18 4 6 10 8 5 8 4 8 6 98 10
826.46 PS(40:10) 6 8 2 20 8 2 2 5 2 2 4 4 4 2 3 5
828.4 PS(40:9) 4 6 2 12 5 2 2 2 2 2 5 4 5 2 3 3
830.5 PS(40:8) 4 4 2 5 3 2 2 2 2 2 5 4 4 2 3 3 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
264
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
832.5 PS(40:7) 4 4 2 2 2 2 2 2 2 2 4 0 4 4 17 3
834.5 PS(40:6) 4 4 2 2 2 2 2 2 2 0 0 0 4 5 12 5
836.5 PS(40:5) 4 4 2 3 2 2 2 2 22 0 0 0 4 5 10 6
838.5 PS(40:4) 4 4 2 3 2 2 2 2 2 0 0 0 4 2 8 6
840.5 PS(40:3) 4 4 2 3 2 2 34 37 31 32 12 12 6 2 8 4
842.5 PS(40:2) 4 4 2 2 2 12 18 16 21 22 10 8 4 6 4 6
844.5 PS(40:1) 4 4 2 2 2 8 8 8 12 18 6 6 4 5 6 4
846.5 PS(40:0) 4 4 2 2 2 4 6 6 4 4 5 4 6 5 7 4
852.48 PS(42:11) 0 0 0 0 0 2 5 5 5 2 3 5 0 0 0 0
854.48 PS(42:10) 0 0 0 0 0 2 4 4 4 2 3 4 0 0 0 0
856.48 PS(42:9) 0 0 0 0 0 2 2 4 4 2 3 4 0 0 0 0
858.48 PS(42:8) 0 0 0 0 0 2 2 4 5 2 3 3 0 0 0 0
860.48 PS(42:7) 0 0 0 0 0 2 15 28 16 22 3 4 0 0 0 0
862.48 PS(42:6) 0 0 0 0 0 2 8 18 14 14 3 3 0 0 0 0
864.48 PS(42:5) 0 0 0 0 0 2 8 15 6 8 3 2 0 0 0 0
866.48 PS(42:4) 0 0 0 0 0 2 6 12 4 6 2 2 0 0 0 0
874.65 PS(42:0) 0 0 0 0 0 2 2 8 2 7 2 2 0 0 0 0
892.6 PS(44:5) 0 0 0 0 0 2 2 5 2 3 8 11 0 0 0 0
765.45 PI(29:1) 4 10 4 8 4 14 15 10 18 14 16 24 10 18 42 20
767.4 PI(29:0) 4 11 5 6 2 2 8 4 8 6 6 10 35 8 21 11
777.4 PI(30:2) 4 12 6 4 17 2 8 4 2 5 4 8 16 0 0 0
791.4 PI(31:2) 0 0 0 0 0 98 90 98 90 81 96 94 8 6 50 51
793.48 PI(31:1) 76 98 98 48 33 30 32 25 40 38 40 20 48 24 68 45
795.5 PI(31:0) 25 36 27 18 16 16 16 12 18 18 20 6 32 34 42 12
799.4 PI(32:5) 6 16 6 6 2 0 0 0 0 0 0 0 8 6 24 8
821.5 PI(33:1) 32 35 22 8 12 0 0 0 0 0 0 0 24 27 30 12
823.5 PI(33:0) 16 18 16 4 8 0 0 0 0 0 0 0 12 15 14 8
825.4 PI(34:6) 6 8 11 22 5 0 0 0 0 0 0 0 8 6 8 7 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
265
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
827.5 PI(34:5) 3 4 2 26 5 0 0 0 0 0 0 0 7 7 7 2
829.4 PI(34:4) 3 4 2 6 2 0 0 0 0 0 0 0 6 7 5 3
831.4 PI(34:3) 2 4 2 4 2 0 0 0 0 0 0 0 6 10 18 8
833.4 PI(34:2) 2 4 2 4 2 0 0 0 0 0 0 0 6 8 16 6
835.4 PI(34:1) 2 4 2 4 2 0 0 0 0 0 0 0 6 6 14 5
837.4 PI(34:0) 2 4 2 5 2 0 0 0 0 0 0 0 5 6 12 4
849.4 PI(36:8) 0 0 0 0 0 0 0 0 0 0 0 0 5 7 10 4
851.4 PI(36:7) 0 0 0 0 0 0 0 0 0 0 0 0 5 5 9 2
853.4 PI(36:6) 2 4 2 5 2 0 0 0 0 0 0 0 4 4 13 2
855.4 PI(36:5) 2 2 4 4 2 0 0 0 0 0 0 0 4 4 14 2
857.4 PI(36:4) 2 4 2 2 2 0 0 0 0 0 0 0 4 4 8 2
859.4 PI(36:3) 2 3 4 2 2 0 0 0 0 0 0 0 4 2 7 2
861.4 PI(36:2) 2 2 2 2 2 4 28 18 24 29 4 4 2 2 4 2
863.8 PI(36:1) 3 3 3 2 2 4 8 6 18 19 4 4 2 2 4 2
865.8 PI(36:0) 2 2 2 2 2 3 6 4 7 8 3 3 2 2 4 2
875.4 PI(38:9) 0 0 0 0 0 0 0 0 0 0 0 0 4 2 12 2
877.4 PI(38:8) 0 0 0 0 0 0 0 0 0 0 0 0 5 10 16 4
879.4 PI(38:7) 0 0 0 0 0 0 0 0 0 0 0 0 4 8 11 4
889.4 PI(38:2) 0 0 0 0 0 3 3 2 10 3 12 12 0 0 0 0
891.4 PI(38:1) 0 0 0 0 0 2 3 2 8 4 8 8 0 0 0 0
893.5 PI(38:0) 0 0 0 0 0 4 3 2 7 3 6 7 0 0 0 0
975.6 PI(44:1) 0 0 0 0 0 2 8 10 20 12 2 2 0 0 0 0
977.6 PI(44:0) 0 0 0 0 0 2 6 8 14 7 2 2 0 0 0 0
535.34 PA(24:0) 5 5 2 4 2 0 0 0 0 0 0 0 0 0 0 0
603.4 PA(29:1) 0 0 0 0 0 2 4 2 8 5 4 4 0 0 0 0
645.4 PA(32:1) 0 0 0 0 0 4 2 2 4 4 4 8 5 5 5 4
687.4 PA(36:8) 2 2 4 2 2 2 2 2 2 4 5 5 4 2 5 4
689.4 PA(36:7) 6 8 3 2 2 4 2 2 18 5 6 9 4 2 3 5 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
266
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
691.4 PA(36:6) 4 6 4 8 2 2 2 2 11 4 12 6 4 2 4 4
693.4 PA(36:5) 5 5 3 6 2 2 2 2 4 4 16 5 3 2 3 3
695.4 PA(36:4) 2 3 3 4 2 2 2 2 3 4 10 4 3 2 4 2
697.4 PA(36:3) 0 0 0 0 0 2 2 2 3 4 6 4 5 2 3 2
699.4 PA(36:2) 0 0 0 0 0 2 2 2 3 3 7 4 5 2 4 2
739.43 PA(40:10) 0 0 0 0 0 0 0 0 0 0 0 0 4 2 5 2
741.4 PA(40:9) 0 0 0 0 0 0 0 0 0 0 0 0 4 4 5 3
713.4 PA(38:9) 0 0 0 0 0 2 2 2 4 2 8 4 0 0 0 0
715.4 PA(38:8) 0 0 0 0 0 2 4 8 5 2 12 5 0 0 0 0
717.4 PA(38:7) 0 0 0 0 0 2 5 6 4 2 40 5 0 0 0 0
719.4 PA(38:6) 0 0 0 0 0 2 4 4 4 2 26 8 0 0 0 0
721.4 PA(38:5) 0 0 0 0 0 2 2 2 4 2 12 7 0 0 0 0
723.4 PA(38:4) 0 0 0 0 0 2 2 2 4 2 11 5 0 0 0 0
725.4 PA(38:3) 0 0 0 0 0 2 2 2 4 2 6 5 0 0 0 0
727.4 PA(38:2) 0 0 0 0 0 2 2 2 4 2 5 5 0 0 0 0
729.4 PA(38:1) 0 0 0 0 0 2 2 2 4 5 60 5 0 0 0 0
731.5 PA(38:0) 2 6 2 2 2 2 3 2 4 3 30 15 0 0 0 0
739.43 PA(40:10) 3 12 18 4 4 2 4 3 4 3 11 12 0 0 0 0
741.4 PA(40:9) 6 12 11 3 18 0 0 0 0 0 0 0 0 0 0 0
743.4 PA(40:8) 6 10 8 2 12 0 0 0 0 0 0 0 4 4 4 4
745.4 PA(40:7) 7 8 4 2 8 0 0 0 0 0 0 0 4 4 4 5
747.4 PA(40:6) 6 7 4 2 4 0 0 0 0 0 0 0 4 4 4 6
749.4 PA(40:5) 0 0 0 0 0 0 0 0 0 0 0 0 4 4 3 4
751.4 PA(40:4) 0 0 0 0 0 0 0 0 0 0 0 0 5 3 3 2
753.4 PA(40:3) 0 0 0 0 0 0 0 0 0 0 0 0 5 3 3 2
755.4 PA(40:2) 0 0 0 0 0 0 0 0 0 0 0 0 5 3 2 2
765.45 PA(42:11) 0 0 0 0 0 15 12 8 22 21 12 20 10 20 28 18
767.45 PA(42:10) 0 0 0 0 0 8 6 5 8 10 6 13 36 22 11 8 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Table continued..
Appendix
267
Mass Compound name UL UF UT UB UZ GC GD GS GTx GFU GFO GCY PT ST CI SA
769.45 PA(42:9) 0 0 0 0 0 6 4 2 7 6 6 7 20 10 8 7
775.45 PA(42:6) 2 7 6 2 2 0 0 0 0 0 0 0 0 0 0 0
777.45 PA(42:5) 2 7 7 2 2 0 0 0 0 0 0 0 0 0 0 0
779.45 PA(42:4) 2 6 6 2 16 0 0 0 0 0 0 0 0 0 0 0
787.6 PA(42:0) 2 4 4 2 2 0 0 0 0 0 0 0 0 0 0 0
813.5 PA(44:1) 17 10 11 4 98 6 14 12 16 16 4 4 24 12 11 7
815.5 PA(44:0) 8 20 8 4 27 4 10 10 12 11 4 4 22 11 14 8 UL-Ulva lactuca, UF-Ulva fasciata, UT-Ulva taeniata, UB- Ulva beytensis, UTb-Ulva tubulosa, GC- Gracilaria corticata, GCY- Gracilaria corticata v. cylindrica, GFO- Gracilaria corticata v. folifera, GD-Gracilaria dura, GS-
Gracilaria salicornia, GTx-Gracilaria textorii, GFU- Gracilaria fergusonii, PT- Padina tetrastomatica, ST- Sargassum tenerrimum, CI- Cystoseira indica, SA- Spatoglossum asperum.
Publications
1. Puja Kumari, A. J. Bijo, Vaibhav A. Mantri, C. R. K Reddy, Bhavanath Jha (2013)
Fatty acid profiling of tropical marine macroalgae: An analysis from chemotaxonomic
and nutritional perspectives. Phytochemistry 86: 44-56. (IF: 3.35, Citations: 1)
2. Puja Kumari, Ravindra Pal Singh, A. J. Bijo, C. R. K. Reddy, Bhavanath Jha (2012)
Estimation of lipid hydroperoxide levels in tropical marine macroalgae. Journal of
Phycology 48: 1362-1373. (IF: 2.07, Citations: 0)
3. Puja Kumari, C. R. K. Reddy, Bhavanath Jha (2011) Comparative evaluation and
selection of a method for lipid and fatty acid extraction from macroalgae. Analytical
Biochemistry 415: 134-144. (IF: 2.99, Citations: 10)
4. Puja Kumari, Manoj Kumar, Vishal Gupta, C. R. K. Reddy, Bhavanath Jha (2010)
Tropical marine macroalgae as potential sources of nutritionally important PUFAs. Food
Chemistry 120: 749-757. (IF: 3.45, Citations: 36)
5. Manoj Kumar, Puja Kumari, Nitin Trivedi, Mahendra Kumar Shukla, Vishal Gupta, C.
R. K. Reddy, Bhavanath Jha (2011). Minerals, PUFAs and antioxidant properties of some
tropical seaweeds from Saurashtra coast of India. Journal of Applied Phycology 23: 797-
810. (IF: 2.4, Citations: 11)
6. Vishal Gupta, Manoj Kumar, Puja Kumari, C. R. K. Reddy, Bhavanath Jha (2011)
Optimization of protoplast yields from the red algae Gracilaria dura (C. Agardh) J.
Agardh and G. verrucosa (Huds.) Papenfuss. Journal of Applied Phycology 23: 209-218.
(IF: 2.4, Citations: 5)
7. Vaibhav A. Mantri, Ravindra Pal Singh, A. J. Bijo, Puja Kumari, C. R. K. Reddy,
Bhavanath Jha (2011) Differential response of varying salinity and temperature on
zoospore induction, regeneration and daily growth rate in Ulva fasciata (Chlorophyta,
Ulvales). Journal of Applied Phycology 23: 243-250. (IF: 2.4, Citations: 3)
8. Manoj Kumar, Vishal Gupta, Nitin Trivedi, Puja Kumari, A. J. Bijo, C. R. K. Reddy,
Bhavanath Jha (2011) Desiccation induced oxidative stress and its biochemical responses
in intertidal red alga Gracilaria corticata (Gracilariales, Rhodophyta). Environmental
and Experimental Botany 72: 194-201. (IF: 2.98, Citations: 4)
9. Ravindra Pal Singh, Mahendra Kumar Shukla, Avinash Mishra, Puja Kumari, C. R. K.
Reddy, Bhavanath Jha (2011) Isolation and characterization of exopolysaccharides from
seaweed associated bacteria Bacillus licheniformis. Carbohydrate Polymers. 84(3): 1019-
1026. (IF: 3.62, Citations: 11)
10. Nitin Trivedi, Vishal Gupta, Manoj Kumar, Puja Kumari, C. R. K. Reddy, Bhavanath
Jha. (2011) An alkali-halotolerant cellulase from Bacillus flexus isolated from green
seaweed Ulva lactuca. Carbohydrate Polymers. 83(2): 891-897. (IF: 3.62, Citations: 9)
11. Nitin Trivedi, Vishal Gupta, Manoj Kumar, Puja Kumari, C. R. K. Reddy, Bhavanath
Jha (2011) Solvent tolerant marine bacterium Bacillus aquimaris secreting organic
solvent stable alkaline cellulose. Chemosphere 83: 706-712. (IF: 3.20; Citations: 3)
12. Vishal Gupta, Ravi S. Baghel, Manoj Kumar, Puja Kumari, Vaibhav A. Mantri, C. R. K.
Reddy, Bhavnath Jha (2011) Growth and agarose characteristics of isomorphic
gametophyte (male and female) and sporophyte of Gracilaria dura and their marker
assisted selection. Aquaculture 318 (3-4): 389-396. (IF: 2.04; Citations: 0)
13. Ravi S. Baghel, Puja Kumari, A. J. Bijo, Vishal Gupta, C. R. K. Reddy, Bhavanath Jha
(2011) Genetic analysis and marker assisted identification of life phases of red alga
Gracilaria corticata (J. Agardh). Molecular Biology Reports 38: 4211-4218. (IF: 2.99;
Citations: 0)
14. Manoj Kumar, Vishal Gupta, Puja Kumari, C. R. K. Reddy, Bhavanath Jha (2010).
Assessment of Caulerpa species for nutritional, fatty acids and antioxidant potentials.
Journal of Food Composition and Analysis. 24: 270-278. (IF: 1.94, Citations: 4)
15. Manoj Kumar, Puja Kumari, Vishal Gupta, P. A. Anisha, C. R. K. Reddy and
Bhavanath Jha (2010). Differential responses to cadmium induced oxidative stress in
marine macroalga Ulva lactuca (Ulvales, Chlorophyta). Biometals 23: 315-325. (IF: 2.3,
Citations: 15)
16. Manoj Kumar, Puja Kumari, Vishal Gupta, C. R. K. Reddy, Bhavanath Jha (2010)
Biochemical responses of red alga Gracilaria corticata (Gracilariales, Rhodophyta) to
salinity induced oxidative stress. Journal of Experimental Marine Biology and Ecology
391: 27-34. (IF: 1.91, Citations: 10)
17. Ravindra Pal Singh, Vishal Gupta, Puja Kumari, Manoj Kumar, C. R. K. Reddy,
Kamalesh Prasad, Bhavanath Jha. (2010) Purification and partial characterization of an
extracellular alginate lyase from Aspergillus oryzae isolated from brown seaweed.
Journal of Applied Phycology. 23: 755-762. (IF: 1.79, Citations: 3)
18. Puja Kumari, C. R. K. Reddy, Bhavanath Jha (2012) Quantification of select
endogenous hydroxy-oxylipins from tropical marine macroalgae. Marine Biotechnology
(Submitted).
Book Chapter
1. Puja Kumari, Manoj Kumar, C. R. K. Reddy, Bhavanath Jha (2013) Algal lipids, fatty
acids and sterols. In Herminia Domínguez ed. Functional ingredients from algae for food
and nutraceuticals. Woodhead Publishing Ltd. UK. In press.
Author's personal copy
Fatty acid profiling of tropical marine macroalgae: An analysisfrom chemotaxonomic and nutritional perspectives
Puja Kumari, A.J. Bijo, Vaibhav A. Mantri, C.R.K. Reddy ⇑, Bhavanath Jha
Discipline of Marine Biotechnology and Ecology, CSIR – Central Salt and Marine Chemicals Research Institute, Bhavnagar, Gujarat 364002, India
a r t i c l e i n f o
Article history:
Received 4 May 2012
Received in revised form 18 October 2012
Available online 17 November 2012
Keywords:
Macroalgae
Lipids
Fatty acids
PUFAs
n6/n3 Ratio
Chemotaxonomy
Endosymbiosis
a b s t r a c t
The lipid and fatty acid (FA) compositions for 100 marine macroalgae were determined and discussed
from the context of chemotaxonomic and nutritional perspectives. In general, the lipid contents in mac-
roalgae were low (2.3–20 mg/g fr. wt.) but with substantially high amounts of nutritionally important
polyunsaturated fatty acids (PUFAs) such as LA, ALA, STA, AA, EPA and DHA, that ranged from 10% to
70% of TFAs. More than 90% of the species showed nutritionally beneficial n6/n3 ratio (0.1:1–3.6:1)
(p 6 0.001). A closer look at the FA data revealed characteristic chemotaxonomic features with C18 PUFAs
(LA, ALA and STA) being higher in Chlorophyta, C20 PUFAs (AA and EPA) in Rhodophyta while Phaeophyta
depicted evenly distribution of C18 and C20 PUFAs. The ability of macroalgae to produce long-chain
PUFAs could be attributed to the coupling of chloroplastic FA desaturase enzyme system from a photo-
synthetic endosymbiont to the FA desaturase/elongase enzyme system of a non-photosynthetic eukary-
otic protist host. Further, the principal component analysis segregated the three macroalgal groups with a
marked distinction of different genera, families and orders, Hierarchical cluster analyses substantiated
the phylogenetic relationships of all orders investigated except for those red algal taxa belonging to Gig-
artinales, Ceramiales, Halymeniales and Rhodymeniales for which increased sampling effort is required
to infer a conclusion. Also, the groups deduced from FA compositions were congruent with the clades
inferred from nuclear and plastid genome sequences. This study further indicates that FA signatures
could be employed as a valid chemotaxonomic tool to differentiate macroalgae at higher taxonomic lev-
els such as family and orders.
Ó 2012 Elsevier Ltd. All rights reserved.
1. Introduction
Benthic marine macroalgae, commonly known as seaweeds are
multicellular photosynthetic organisms with considerable poten-
tials for using as a source of bioactive compounds of immense
pharmaceutical and nutraceutical importance. They are rich
sources of nutritionally beneficial components such as proteins,
carbohydrates, polyunsaturated fatty acids (PUFAs), antioxidants,
minerals, dietary fibers and vitamins (Chandini et al., 2008;
Mohamed et al., 2011) and are thus consumed as functional foods.
There are 250 macroalgal species commercially utilized world-
wide, of which 150 are consumed as human food (Barrow, 2007).
The macroalgal species, in general, are low in lipids and contain
1–5% on dry wt. basis. Nevertheless, the nutritionally important
C18 and C20 PUFAs including n3 PUFAs are present in substantially
high amounts with anti-inflammatory, anti-thrombotic and anti-
arrhythmic responses (Kumari et al., 2010; Gillies et al., 2011).
The n-3 PUFAs are of particular importance as they cannot be
synthesized by humans and are thus obtained only through dietary
sources.
Fatty acids (FAs) being metabolites of conserved acetyl-CoA
pathway have been extensively studied from the context of che-
motaxonomic perspectives in higher plants (Mongrand et al.,
2001, 2005; Dussert et al., 2008), cyanobacteria (Shukla et al.,
2011), bacteria (Malviya et al., 2011; Núñez-Cardona, 2012), mic-
roalgae (Dunstan et al., 2005; Lang et al., 2011) and fungi (Mishra
et al., 2010). Further, Dunstan et al. (2005) deciphered the evolu-
tionary relationship between the FA composition of Rhodo-
phyceaen and Cryptophyceaen microalgae and the endosymbiotic
theory. According to the endosymbiotic theory, the micro- and
macroalgae of both Chlorophyceae and Rhodophyceae have been
originated from primary endosymbiosis of photosynthetic cyano-
bacteria and eukaryotic host while Phaeophyta diverged from red
algae via secondary (or tertiary) endosymbiosis along with other
chlorophyll c bearing algae such as Cryptophytes, Haptophytes,
diatoms, Dinoflagellates and non-photosynthetic Apicomplexans,
ciliates and oomycetes, forming the super-group Stramenopiles
at the base of the tree of life (Baldauf et al., 2000; Baldauf, 2008;
Archibald, 2009, 2012; Dorrell and Smith, 2011; Baurain et al.,
2012; De Clerck et al., 2012; Green, 2010; Woehle et al., 2012).
0031-9422/$ - see front matter Ó 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.phytochem.2012.10.015
⇑ Corresponding author. Tel.: +91 278 256 5801x614; fax: +91 278 2567562/
2566970.
E-mail address: [email protected] (C.R.K. Reddy).
Phytochemistry 86 (2013) 44–56
Contents lists available at SciVerse ScienceDirect
Phytochemistry
journal homepage: www.elsevier .com/locate /phytochem
ESTIMATION OF LIPID HYDROPEROXIDE LEVELS IN TROPICALMARINE MACROALGAE1
Puja Kumari, Ravindra Pal Singh, A. J. Bijo, C. R. K. Reddy,2 and Bhavanath Jha
Discipline of Marine Biotechnology and Ecology, CSIR-Central Salt and Marine Chemicals Research Institute, Bhavnagar, 364002,
Gujarat, India
The incipient levels of lipid hydroperoxides(LHPOs) were determined in selected green, brown,and red macroalgae by the FOX assay usinghydroperoxy HPLC mix. The LHPOs contents variedbetween the investigated species and showedrelatively low values in this study. Among the greens,it varied from 12 ± 6.2 lg g!1 (Codium sursum) to31.5 ± 2.8 lg g!1 (Ulva lactuca), whereas in reds,from 5.7 ± 1.6 lg g!1 (Gracilaria corticata) to46.2 ± 6 lg g!1 (Sarconema filiforme), and in browns,from 4.6 ± 4.4 lg g!1 (Dictyota bartayresiana) to79 ± 5.0 lg g!1 (Sargassum tenerrimum), on freshweight basis. These hydroperoxides represented aminor fraction of total lipids and ranged from 0.04%(S. swartzii) to 1.1% (S. tenerrimum) despite being arich source of highly unsaturated fatty acids. Thesusceptibility of peroxidation was assessed by specificlipid peroxidazibility (SLP) values for macroalgaltissues. The LHPO values were found to beindependent of both the PUFAs contents and theirdegree of unsaturation (DBI), as evident from thePCA analysis. SLP values were positively correlatedwith the LHPOs and negatively with DBI. The FOXassay gave " 20-fold higher values for LHPOs ascompared to the TBARS method for all the samplesinvestigated in this study. Furthermore, U. lactucacultured in artificial seawater (ASW) enriched withnutrients (N, P, and NP) showed a sharp decline inLHPOs contents relative to those cultured in ASWalone P # 0.05. It is inferred from this study that theFOX assay is an efficient, rapid, sensitive, andinexpensive technique for detecting the incipientlipid peroxidation in macroalgal tissues.
Key index words: fatty acid; FOX; lipid hydroperoxides;macroalgae; TBARS
Abbreviations: 12(S)-HpETE, 12(S)-Hydroperoxyeico-satetraenoic acid; 15(S)-HpETE, 15(S)-Hydroperoxy-eicosatetraenoic acid; 5(S)-HpETE, 5(S)-Hydroperoxyeicosatetraenoic acid; 13(S)-HpODE, 13(S)-Hydro-peroxyoctadecadienoic acid; 9(S)-HpODE, 9(S)-Hy-droperoxyoctadecadienoic acid; 13-HpOTE, 13-Hydroperoxyoctadecatrienoic acid; ANOVA, analysis ofvariance; ASW, artificial seawater; BHT, butylatedhydroxytoluene; CAT, catalase; DBI, double bond
index; FA, fatty acid; FAME, fatty acid methyl ester;FOX, ferrous oxidation-xylenol orange; FOX1, fer-rous oxidation-xylenol orange version 1; FOX2, fer-rous oxidation-xylenol orange version 2; H2O2,hydrogen peroxide; HPO, hydroperoxy HPLC mix;GC, gas chromatography; GC–MS, gas chromatographymass spectrometry; LC–MS, liquid chromatographymass spectrometry; LHPOs, lipid hydrop-eroxides; MDA, malondialdehyde; PC1, principalcomponent 1; PC2, principal component 2; PCA,principal component analysis; POD, peroxidase;PUFA, polyunsaturated fatty acid; ROS, reactiveoxygen species; SLP, specific lipid peroxidazibility;SOD, superoxide dismutase; TBARS, thiobarbituricacid-reactive substances; TFA, total fatty acid; TL,total lipid; TPP, triphenylphosphine
Lipid peroxidation is a well recognized indicatorof oxidative stress in all types of organisms, includ-ing plants, bacteria, fungi, algae, and mammaliansystems. In addition, their products exhibit a widevariety of biological and cell signaling functions(Spickett et al. 2010). Lipid peroxidation can beeither enzymatic mediated by lipoxygenases andcyclooxygenases, or nonenzymatic free radicalmediated, where several chain reactions are medi-ated by reactive oxygen species (ROS), whichthemselves are unavoidable consequences of aerobicmetabolism.Although lipid peroxidation is a well-investigated
phenomenon, lipid hydroperoxides (LHPOs) havereceived comparatively less attention than the otherlipid oxidized products because of their either insta-bility or lengthy procedures involving of expensiveand sophisticated equipments for detection. TheLHPOs have been estimated by several analyticalapproaches, such as high-performance liquidchromatography (HPLC; Nakamura and Maeda1991, Hui et al. 2005), gas chromatography (GC; Tur-nipseed et al. 1993), electrospray mass spectrometry(Spickett et al. 1998), iodine oxidation (Jessup et al.1994), heme degradation of peroxides (Frei et al.1998), cyclooxygenase activation (Pendleton andLands 1987, Calzada et al. 1997), chemiluminescencedetection (Yamamoto 1994), immunological assays(Paradis et al. 1997), conjugated diene measurement(Moore and Roberts 1998), and thiobarbituric
1Received 29 February 2012. Accepted 5 June 2012.2Author for correspondence: e-mail: [email protected].
J. Phycol. 48, 1362–1373 (2012)
© 2012 Phycological Society of America
DOI: 10.1111/j.1529-8817.2012.01208.x
1362
Author's personal copy
Comparative evaluation and selection of a method for lipid and fatty acidextraction from macroalgae
Puja Kumari, C.R.K. Reddy ⇑, Bhavanath Jha
Discipline of Marine Biotechnology and Ecology, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research (CSIR), Bhavnagar 364021,
Gujarat, India
a r t i c l e i n f o
Article history:
Received 27 January 2011
Received in revised form 16 March 2011
Accepted 6 April 2011
Available online 16 April 2011
Keywords:
Lipids
Fatty acids
Macroalgae
Sonication
Buffer
Extraction method
a b s t r a c t
A comparative evaluation of Bligh and Dyer, Folch, and Cequier-Sánchez methods for quantitative deter-
mination of total lipids (TLs) and fatty acids (FAs) was accomplished in selective green (Ulva fasciata), red
(Gracilaria corticata), and brown algae (Sargassum tenerrimum) using a full factorial categorical design.
Applications of sonication and buffer individually on lipid extraction solvent systems were also evalu-
ated. The FA recoveries obtained from the aforementioned methods were compared with those of direct
transesterification (DT) methods to identify the best extraction methods. The experimental design
showed that macroalgal matrix, extraction method, and buffer were key determinants for TL and FA
recoveries (P 6 0.05), exhibiting significant interactions. But sonication gave erratic results with no inter-
action with any of the factors investigated. The buffered solvent system of Folch rendered the highest TL
yield in U. fasciata and G. corticatawhile the buffered system of Bligh and Dyer gave the highest yield in S.
tenerrimum. DT methods were more convenient and accurate for FA quantification and rendered 1.5–2
times higher yields when compared with the best conventional method, minimizing the use of chlori-
nated solvents, their cost of analysis, and disposal. The buffered solvent system was found to be the most
appropriate for lipid research in macroalgae.
Ó 2011 Elsevier Inc. All rights reserved.
Macroalgae have been reported to contain more than 2400 nat-
ural products of commercial importance in pharmaceutical, bio-
medical, and nutraceutical industries [1]. They have also been
extensively utilized as ingredients in human and animal food prep-
arations owing to their high contents of polyunsaturated fatty
acids (PUFAs),1 carbohydrates, vitamins, minerals, and dietary fibers
[2,3]. Nowadays, algal resources have been studied with renewed
interest across the world as an alternative source of renewable en-
ergy feedstock that circumvents the controversy of ‘‘fuel versus
food’’. The attributes for such choice are their relatively higher pro-
duction turnover and amenability for depolymerization of substrate
in addition to greater carbon sequestration potentials than terrestrial
feedstock [4]. Most recently, Petcavich succeeded in producing
hydrocarbon biofuels from transformed kelp Macrocystis pyrifera
with high hydrocarbon producing genes of microalgae, Botryococcus
braunii [5].
Fatty acid (FA) analysis has been increasingly gaining impor-
tance due to the realization of their beneficial applications in nutri-
tional and health products. Further, they have also been used for
addressing various fundamental and pragmatic research problems
in experimental biochemical, physiological, and clinical studies
[6,7]. Further, in biodiesel production, clean burn properties of
the fuel are influenced by FA structural features including chain
length and degree of unsaturation [8]. Thus, a precise quantifica-
tion of FA can also be used to predict the quality of biodiesel, which
is reduced considerably with the increase in the amount of satu-
rated FAs.
Traditionally, the fatty acid composition of lipid samples is
determined by assessing the corresponding methyl esters via gas
chromatography (GC). A large number of analytical approaches
based on initial lipid extraction by solvents, followed by their
transmethylation (i.e., conventional methods), are employed and
where FAs are sought, they are extracted and methylated with
one-step procedures wherein methylation reagent is added di-
rectly to the samples without previous extraction (direct transeste-
rification methods). However, both types of methods have their
own advantages and disadvantages that are well illustrated in
the literature [9–16]. Conventional methods are time consuming
0003-2697/$ - see front matter Ó 2011 Elsevier Inc. All rights reserved.
doi:10.1016/j.ab.2011.04.010
⇑ Corresponding author. Fax: +91 278 2567562/2566970.
E-mail address: [email protected] (C.R.K. Reddy).1 Abbreviations used: BD, Bligh and Dyer; BDB, Bligh and Dyer with buffer; BDS,
Bligh and Dyer with sonication; CM, conventional methods; CS, Cequier-Sánchez
method; CSB, Cequier-Sánchez method with buffer; CSS, Cequier-Sánchez method
with sonication; DT, direct transesterification; FA, fatty acid; FAMEs, fatty acid methyl
esters; FM, Folch method; FMB, Folch method with buffer; FMS, Folch method with
sonication; GM, Garcia method; LRC, Lepage and Roy modified by Cohen method;
PUFAs, polyunsaturated fatty acids; TFA, total fatty acid; MUFA, monounsaturated
fatty acids; SFA, saturated fatty acid.
Analytical Biochemistry 415 (2011) 134–144
Contents lists available at ScienceDirect
Analytical Biochemistry
journal homepage: www.elsevier .com/locate /yabio
Author's personal copy
Tropical marine macroalgae as potential sources of nutritionally important PUFAs
Puja Kumari, Manoj Kumar, Vishal Gupta, C.R.K. Reddy *, Bhavanath Jha *
Discipline of Marine Biotechnology and Ecology, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research (CSIR), Bhavnagar 364021, India
a r t i c l e i n f o
Article history:
Received 27 January 2009
Received in revised form 25 September
2009
Accepted 4 November 2009
Keywords:
Chlorophyta
Fatty acids
Lipids
n6/n3 ratio
Phaeophyta
PUFAs
Rhodophyta
Tropical macroalgae
a b s t r a c t
The lipid and fatty acid compositions of 27 tropical macroalgae belonging to the three phyla, Chlorophyta,
Phaeophyta and Rhodophyta, were studied from a nutritional and chemotaxonomic perspective. The lipid
content varied widely among the species and ranged from 0.57% to 3.5% on a dry weight basis (p 6 0.01).
Chlorophyta members showed higher C18PUFAs contents than did C20 PUFAs while for Rhodophyta the
trend was opposite. The Phaeophyta members displayed a profile of C18PUFAs similar to that of Chloro-
phyta and of C20PUFAs to that of Rhodophyta. Both Phaeophyta and Rhodophyta species were rich in
arachadonic acid (AA) and eicosopentaenoic acid (EPA) and Ulvales in docosahexaenoic acid (DHA) con-
tent. Most of the species studied had a nutritionally beneficial n6/n3 ratio (0.61–5.15:1). Further, the
principal component analysis clearly segregated the three phyla by their FA composition and hierarchical
cluster analysis altogether classified them into six distinct groups, suggesting that FAs can be used as a
tool for chemotaxonomic studies.
Ó 2009 Elsevier Ltd. All rights reserved.
1. Introduction
Benthic marine macroalgae, commonly known as seaweeds,
are increasingly viewed as potential sources of bioactive com-
pounds with immense pharmaceutical, biomedical and nutraceu-
tical importance (Veena, Josephine, Preetha, & Varalakshmi,
2006). Many macroalgal species have been used as ingredients
in both medicinal and food preparations, traditionally, in different
regions across the world (Chandini, Ganesan, Suresh, & Bhaskar,
2008). In addition, some are common sources of phycocolloids
(gelling agents) of commercial value (Cardozo et al., 2007). There
are 250 macroalgal species which have been listed as commer-
cially utilised worldwide, among which 150 are consumed as
human food (Barrow, 2007). They are also considered as low cal-
orie foods with high contents of minerals, vitamins, proteins and
carbohydrates (Chandini et al., 2008). Although macroalgae have
been reported to have low lipid contents, their polyunsaturated
fatty acid (PUFA) contents are superior to those of the terrestrial
vegetables (Darcy-Vrillon, 1993). They are rich in C18 and C20
PUFAs with nutritional implications and are thus, studied
extensively for biotechnological, food, feed, cosmetic and pharma-
ceutical applications (Chandini et al., 2008). Long-chain n 3 PU-
FAs, such as EPA and DHA, have various beneficial clinical and
nutraceutical applications (Ginzberg et al., 2000; Lombardo, Hein,
& Chicco, 2007). Their role in growth, development and reproduc-
tion of both marine invertebrates and fish has also been well
documented (Rodriguez et al., 2004). The n 3 PUFAs cannot be
synthesized by humans and are thus obtained through diet. In
view of their promising medical and nutritional applications, they
have been extensively investigated, However, the studies on effi-
cient exploitation of natural sources for these compounds are lim-
ited (Colombo et al., 2006). At present, marine fishes and fish oils
are the main commercial sources of PUFAs but their suitability for
human consumption has been questioned from a biosafety
perspective, raising the need to search for alternative sources of
high quality PUFAs (Bhosale, Velankar, & Chaugule, 2008). Conse-
quently, marine macroalgae have been studied as alternative
potential sources, as many of them could easily be cultivated in
the sea on a large scale (Critchely, Ohno, & Largo, 2006). Also,
the PUFAs present in the fishes enter the food chain from differ-
ent trophic levels as a result of consuming primary producers,
such as phytoplankton and seaweeds, which synthesize and store
them in good quantities (Visentainer et al., 2007).
Further, FAs have been extensively used as a chemotaxonomic
tool to classify the species in higher plants (Mongrand et al.,
2001) and cyanobacteria (Temina, Rezankovab, Rezankac, &
Dembitsky, 2007). Recently, Marsham, Scott, and Tobin (2007)
grouped seaweeds by their nutritional composition using multi-
variate analysis. To the best of our knowledge, classification of
macroalgae by FAs remains largely unexplored to date.
0308-8146/$ - see front matter Ó 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2009.11.006
* Corresponding authors. Tel.: +91 278 256 7352; fax: +91 278 257 0885/256
6970/256 7562.
E-mail address: [email protected] (B. Jha).
Food Chemistry 120 (2010) 749–757
Contents lists available at ScienceDirect
Food Chemistry
journal homepage: www.elsevier .com/locate / foodchem
Minerals, PUFAs and antioxidant properties of some
tropical seaweeds from Saurashtra coast of India
Manoj Kumar & Puja Kumari & Nitin Trivedi &
Mahendra K. Shukla & Vishal Gupta & C. R. K. Reddy &
Bhavanath Jha
Received: 16 April 2010 /Revised and accepted: 17 August 2010 /Published online: 7 September 2010# Springer Science+Business Media B.V. 2010
Abstract Twenty-two tropical seaweeds from the Rhodo-
phyta, Phaeophyta and Chlorophyta were examined for
their possible use as nutritional supplements. All seaweeds
contained balanced Na/K and C/N ratio and high amounts
of macroelements (Na, K, Ca, and Mg) as compared to the
terrestrial vegetables. Among the microelements, Fe was
the highest followed by Zn, Mn, Cu and other trace
elements. Fatty acid distribution showed high level of n-6
and n-3 polyunsaturated fatty acids (PUFAs), and their
ratios were within the WHO prescribed limits. The higher
ratios of PUFA/SFA (>0.4) are in agreement with the
recommendations of nutritional guidelines. Most of the
species, especially the Chlorophyta and Phaeophyta, had
permissible intake values of unsaturation, atherogenic and
thrombogenic indexes comparable to milk-based products.
Principal component analysis demonstrated a correlation
between total phenolic content, total antioxidant activity,
DPPH, and O2•− radical scavenging activity, suggesting
polyphenols as the chief contributor to the antioxidant
activity in seaweeds. These results indicate that these
seaweeds could be a potential source of natural antiox-
idants, minerals and high-quality PUFAs and may be
efficiently used as ingredients in functional foods.
Keywords Antioxidant potential . Minerals . PUFAs .
Tropical seaweeds
Introduction
Increasing awareness among consumers about health-
promoting foods has aroused interest in food supplement
research worldwide. In addition to food supplements,
consumption of exotic foods with proven nutritional
values has also been gaining prominence in several
developed countries (Herrero et al. 2006). Many of these
foods are presently promoted and marketed as function-
al foods with premium price. The beneficial actions of
these foods are reported to be mainly due to their function-
al components such as minerals, antioxidants and n-3 fatty
acids, which are either absent in the analogous traditional
foods or present only in trace concentrations. Consequent-
ly, there has been a quest to explore and utilize foods from
nonconventional sources of both terrestrial and marine
origin to enhance the nutritional quality of human foods
which in turn also reduces the dependability on traditional
foods.
Seaweeds with their diverse bioactive compounds (Lee
et al. 2008; Zubia et al. 2009) have opened up potential
opportunities in pharmaceutical and agri-food processing
industries. The consumption of seaweeds as part of diet has
been shown to be one of the prime reasons for low
incidence of breast and prostate cancer in Japan and China
compared to North America and Europe (Pisani et al.
2002). Seaweeds also contain sufficient amounts of protein,
polysaccharides (e.g., alginates, fucans and laminarans),
and amino acids of considerable nutritional importance.
Algal lipids (1–3% dry matter) contain a high proportion of
essential fatty acids particularly n-3 polyunsaturated fatty
acids (PUFAs). At present, marine fish and their oil are the
major commercial sources of PUFAs, but their suitability
for human consumption has been questioned from the
biosafety perspective.
M. Kumar : P. Kumari :N. Trivedi :M. K. Shukla :V. Gupta :
C. R. K. Reddy (*) : B. Jha
Discipline of Marine Biotechnology and Ecology,
Central Salt and Marine Chemicals Research Institute,
Council of Scientific and Industrial Research (CSIR),
G.B. Marg,
Bhavnagar 364021, India
e-mail: [email protected]
J Appl Phycol (2011) 23:797–810
DOI 10.1007/s10811-010-9578-7
Optimization of protoplast yields from the red algae
Gracilaria dura (C. Agardh) J. Agardh and G. verrucosa
(Huds.) Papenfuss
Vishal Gupta & Manoj Kumar & Puja Kumari &
C. R. K. Reddy & Bhavanath Jha
Received: 4 February 2010 /Revised and accepted: 17 August 2010# Springer Science+Business Media B.V. 2010
Abstract This study reports on the optimization of
protoplast yield from two important tropical agarophytes
Gracilaria dura and Gracilaria verrucosa using different
cell-wall-degrading enzymes obtained from commercial
sources. The conditions for achieving the highest protoplast
yield was investigated by optimizing key parameters such
as enzyme combinations and their concentrations, duration
of enzyme treatment, enzyme pH, mannitol concentration,
and temperature. The significance of each key parameter
was also further validated using the statistical central
composite design. The enzyme composition with 4%
cellulase Onozuka R-10, 2% macerozyme R-10, 0.5%
pectolyase, and 100 U agarase, 0.4 M mannitol in seawater
(30‰) adjusted to pH 7.5 produced the highest protoplast
yields of 3.7±0.7×106 cells g−1 fresh wt for G. dura and
1.2±0.78×106 cells g−1 fresh wt for G. verrucosa when
incubated at 25°C for 4–6 h duration. The young growing
tips maximally released the protoplasts having a size of 7–
15 μm in G. dura and 15–25 μm in G. verrucosa, mostly
from epidermal and upper cortical regions. A few large-size
protoplasts of 25–35 μm, presumably from cortical region,
were also observed in G. verrucosa.
Keywords Agarophytes . Commercial enzymes .
Gracilaria . Protoplast production . Rhodophyta
Introduction
Among the red algae, the genus Gracilaria has considerable
industrial importance as an agarophyte and is the principal
source of raw material to the agar industry worldwide
(Zemke-White and Ohno 1999; Smit 2004). These agar
industries consume 72,300 dry tons of agarophytes annually
and produce approximately 9,600 tons agar with a value
of US$173 million. Of this, Gracilaria alone accounts for
about 80% of the world’s agar market with a value of
US$ 138 million (Bixler and Porse 2010). The increasing
demand for agar worldwide, coupled with short supplies of
agarophytes from wild stocks, has led to the development
of viable field cultivation methods for their large-scale
cultivation in the sea (Critchley 1993). Following the success
in large-scale cultivation, cellular biotechnology techniques
are also being applied to improve the cultivated germplasm
of this important resource (see reviews of Reddy et al.
2008a, b, 2010). However, these techniques have largely
remained underdeveloped and are thus in their nascent stage.
The high structural complexity and diversity in cell wall
composition (Duckworth and Yaphe 1971; Rochas and
Lahaye 1989) have rendered the agarophytes in particular
recalcitrant to enzymatic digestion of the cell walls and have
thus become an impediment for realizing the potentials
offered by the application of biotechnology tools and
techniques for seaweeds in general. The skeletal and matrix
polysaccharides of cell walls of red seaweeds mainly consist
of cellulose and agar. Therefore, the mixture of enzymes
having cellulase and agarase are invariably required to digest
the cell wall components of agarophytic red algal species for
preparing protoplasts.
To date, protoplast isolation has been accomplished for
48 red algal species, including agarophytes, belonging to 13
genera (Reddy et al. 2010). In the genus Gracilaria, with
This paper was presented at the 7th Asia Pacific Congress on Algal
Biotechnology, New Delhi, 2009.
V. Gupta :M. Kumar : P. Kumari :C. R. K. Reddy (*) :B. Jha
Discipline of Marine Biotechnology and Ecology,
Central Salt and Marine Chemicals Research Institute,
Council of Scientific and Industrial Research (CSIR),
Bhavnagar 364021, India
e-mail: [email protected]
J Appl Phycol
DOI 10.1007/s10811-010-9579-6
Differential response of varying salinity and temperature
on zoospore induction, regeneration and daily growth rate
in Ulva fasciata (Chlorophyta, Ulvales)
Vaibhav A. Mantri & Ravindra Pal Singh & A. J. Bijo &
Puja Kumari & C. R. K. Reddy & Bhavanath Jha
Received: 17 January 2010 /Revised and accepted: 11 June 2010# Springer Science+Business Media B.V. 2010
Abstract Seaweed cultivation is imperative to augment
increasing industrial demand. Ulva fasciata Delile is a
potential seaweed for cultivation with applications in food
industries. There is a renewed interest in large-scale
aquaculture of this species in India due to its envisaged
demand in snack food products. In the present study, we
have successfully demonstrated the possibility of inducing
zoospores in vegetative tissue, effective regeneration and
improved growth in this seaweed by manipulating salinity
(from 15 to 30 psu) and temperature (from 15 to 35°C). The
optimum salinity and temperature requirement for zoo-
spores induction were found to be 15 psu and 25°C,
respectively. The quadriflagellate zoospores showed nega-
tive phototaxis and the settlement and germination pattern
similar to several other green seaweeds. The optimum
regeneration (78.53±10.05%) was recorded at 25°C and
30 psu salinity. The maximum daily growth rate (16.1±
0.28%) was at 25°C and 30 psu salinity which corre-
sponded to the field conditions. This method could be
further refined at nursery culture to achieve artificial
seeding essential for the success of commercial cultivation
of this seaweed.
Keywords Cultivation . Edible seaweed . Salinity .
Temperature .Ulva fasciata
Introduction
The green alga Ulva fasciata Delile (Chlorophyta, Ulvales)
is a potential edible seaweed distributed widely along the
Northern west coast of India. This seaweed is a rich source
of proteins, essential amino acids, fatty acids, vitamins,
dietary fibers and minerals vital for human nutrition (Sitaka
Rao and Tipnis 1964; Lewis 1966; McDermid and Stuercke
2003; Carvalho et al. 2009; Kumari et al. 2010). The iodine
content has been estimated in a range of 29–37 mg per
100 g dry wt. (Kesava Rao and Indusekhar 1989). A
substantial amount of beta-carotene and alpha tocopherol
has also been reported (de Sousa et al. 2008). The naturally
harvested biomass from the West coast of India has been
reported to possess antiviral properties against Japanese
encephalitis virus (Sharma et al. 1996). Recently, local food
industries have developed interest in incorporating this
species in popular snack items. Its dark green color, rich
flavor, and sweet aroma along with wide range of
nutritionally beneficial substances including omega-3-fatty
acids can improve dietetic composition of the food items.
This suggests a potential high demand for this species by
the food processing sector. However, inadequate natural
resources may hamper its prospects for utilization by food
industry. The development of reliable cultivation technique
based on the artificial seed production can ensure sustain-
able supply of quality biomass thereby negating the
consequences of over-harvesting.
Species of Monostroma, Enteromorpha, and Ulva are
popular in Japanese cuisine and are being cultivated
commercially in Japan and other Southeast Asian countries
(Ohno 2006; Hiraoka and Oka 2008). The seedling
production required for large-scale cultivation of these
seaweeds has traditionally been achieved through entrap-
ment of the zoospores on cultivation nets kept under the
This paper was presented at the 7th Asia Pacific Congress on Algal
Biotechnology, New Delhi 2009.
V. A. Mantri (*) : R. P. Singh : A. J. Bijo : P. Kumari :
C. R. K. Reddy : B. Jha
Marine Biotechnology and Ecology Discipline,
Central Salt and Marine Chemicals Research Institute,
Council of Scientific and Industrial Research (CSIR),
Gijubhai Badheka Marg,
Bhavnagar 364 021, India
e-mail: [email protected]
J Appl Phycol
DOI 10.1007/s10811-010-9544-4
Environmental and Experimental Botany 72 (2011) 194–201
Contents lists available at ScienceDirect
Environmental and Experimental Botany
journa l homepage: www.e lsev ier .com/ locate /envexpbot
Desiccation induced oxidative stress and its biochemical responses in intertidal
red alga Gracilaria corticata (Gracilariales, Rhodophyta)
Manoj Kumar, Vishal Gupta, Nitin Trivedi, Puja Kumari, A.J. Bijo, C.R.K. Reddy ∗, Bhavanath Jha
Discipline of Marine Biotechnology and Ecology, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research (CSIR), Bhavnagar 364021, India
a r t i c l e i n f o
Article history:
Received 9 July 2010
Received in revised form 4 March 2011
Accepted 11 March 2011
Keywords:
Antioxidative enzymes
Desiccation
Gracilaria corticata
Polyamines
PUFAs
Reactive oxygen species
a b s t r a c t
Intertidal alga Gracilaria corticata growing in natural environment experiences various abiotic stresses
during the low tides. The aim of this study was to determine whether desiccation exposure would lead
to oxidative stress and its effect varies with exposure periods. This study gives an account of various
biochemical changes in G. corticata following the exposure to desiccation for a period of 0 (control), 1,
2, 3 and 4 h under controlled conditions. During desiccation, G. corticata thalli showed dramatic loss
of water by almost 47% when desiccated for 4 h. The enhanced production of reactive oxygen species
(ROS) and increased lipid peroxidation observed during the exposure of 3–4 h were chiefly contributed
by higher lipoxygenase (LOX) activity with the induction of two new LOX isoforms (LOX2, ∼85 kDa;
LOX3, ∼65 kDa). The chlorophyll, carotenoids and phycobiliproteins (phycoerythrin and phycocyanin)
were increased during initial 2 h exposure compared to control and thereafter declined in the succeeding
exposure. The antioxidative enzymes such as superoxide dismutase (SOD), ascorbate peroxidase (APX),
glutathione reductase (GR), glutathione peroxidase (GPX) and the regeneration rate of reduced ascorbate
(AsA) and glutathione (GSH) increased during desiccation up to 2–3 h. Further, the isoforms of antiox
idant enzymes MnSOD (∼150 kDa), APX4 (∼110 kDa), APX5 (∼45 kDa), GPX1 (∼80 kDa) and GPX2
(∼65 kDa) responded specifically to the desiccation exposure. Compared to control, a relative higher con
tent of both free and bound insoluble putrescine and spermine together with enhanced n6 PUFAs namely
C20:4(n6) and C20:3(n6) fatty acids found during 2 h exposure reveals their involvement in defence
reactions against the desiccation induced oxidative stress.
© 2011 Elsevier B.V. All rights reserved.
1. Introduction
The red alga Gracilaria corticata (J. Agardh) J. Agardh occurs
extensively in intertidal zone of the Indian coast and regularly
experiences the desiccation during low tide periods. The organ
isms living in the intertidal zone of tropical shores are subjected
to various types of abiotic stresses due to periodic exposure to
a wide range of fluctuating environmental factors such as desic
cation, salinity, radiation, temperature and pollutants (Apel and
Hirt, 2004; Liu and Pang, 2010; Kumar et al., 2010a). The environ
mental exposure during low tide condition, demands the intertidal
macroalgae to prepare early for the desiccation followed by rehy
dration and associated cellular damage (Burritt et al., 2002). This
constant state of readiness requires a great deal of energy budget
and could be a contributing factor to the slow growth rates of algae
dwelling at the upper littoral zone as compared to those at lower
littoral zone (Stengal and Dring, 1997). The possible explanation
∗ Corresponding author. Tel.: +91 278 256 5801/3805x614;
fax: +91 278 256 6970/7562.
Email address: [email protected] (C.R.K. Reddy).
for the success of an alga exposed to drought could be either being
physiologically more tolerant or better at resisting the water loss
(Ji and Tanaka, 2002).
During desiccation many of the intertidal seaweeds experience
extreme drying rates, reaching air dryness within hours (Schonbeck
and Norton, 1979; Nelson et al., 2010), generally depends on the cli
matic conditions as well as the evaporating surfacetovolume ratio
of the thallus (Lobban et al., 1985). Also, desiccation causes cellu
lar dehydration, which increases the concentration of electrolyte
within the cell, causing changes to membranebound structures
including the thylakoid (Kim and Garbary, 2007). It has been sug
gested that instant responses of marine plants to adverse milieu
involve excess production of reactive oxygen species (ROS) such
as hydrogen peroxide (H2O2), singlet oxygen (1O2), superoxide
(O2•−) and hydroxyl radical (OH−) (Burritt et al., 2002). The abil
ity to withstand the oxidative assault imposed by ROS depends
on the enzymatic and non enzymatic oxidants of the cell. This
antioxidant system functions in a coordinated manner to alle
viate the cellular hypo/hyper osmolarity, ion disequilibrium and
detoxification of ROS which otherwise cause oxidative destruc
tion to cell (Wu and Lee, 2008; Liu and Pang, 2010; Kumar et al.,
2010a,b).
00988472/$ – see front matter © 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.envexpbot.2011.03.007
Carbohydrate Polymers 84 (2011) 1019–1026
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Isolation and characterization of exopolysaccharides from seaweed associated
bacteria Bacillus licheniformis
Ravindra Pal Singh, Mahendra K. Shukla, Avinash Mishra, Puja Kumari, C.R.K. Reddy ∗, Bhavanath Jha
Discipline of Marine Biotechnology and Ecology, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research (CSIR), Bhavnagar 364021, India
a r t i c l e i n f o
Article history:
Received 2 December 2010
Received in revised form
17 December 2010
Accepted 19 December 2010
Available online 25 December 2010
Keywords:
Exopolysaccharide
MALDITOFTOF MS
Bacillus licheniformis
Seaweed
Atomic force microscopy
Xray diffraction
a b s t r a c t
In the present study, EPS secreted by the endophytic bacterium Bacillus licheniformis was isolated and
characterized. The molecular masses of the EPS were 1540 and 44,565 kDa and 1H NMR, FTIR and UV–Vis
spectral analyses revealed prevalence of characteristic primary aminegroup, aromaticcompound, halide
and aliphatic alkylgroup in addition to Na, P, Ca, C, O, Cl and S as inferred from EDX analysis. XRD and DSC
analysis confirmed the amorphous nature of EPS, showing an average particle size of 24.977 mm (d 0.5)
with 191 nm average roughness. The positive ion reflector mode of MALDI TOFTOF MS exhibited a series
of low and high mass peaks corresponding to various oligosaccharides and polysaccharides respectively.
Further, GC–MS analysis revealed its four monosaccharide constituents glucose, galactose, mannose and
arabinose. The potential heterogeneous properties of EPS as revealed in the present study may be explored
in various biotechnological and industrial applications.
© 2010 Elsevier Ltd. All rights reserved.
1. Introduction
Microbial exopolysaccharides (EPSs) are biosynthetic polymers
mainly consisting of carbohydrates secreted by bacteria (Freitas
et al., 2009) and cyanobacteria (Chi, Su, & Lu, 2007; Decho,
1990; Parikh & Madamwar, 2006), however, it is also reported
from Cryptophyta (Bermudez, Rosales, Loreto, Briceno, & Morales,
2004), mushroom (Zou, Sun, & Guo, 2006), yeast (Duan, Chi,
Wang, & Wang, 2008) and basidiomycetes (Chi & Zhao, 2003;
Manzoni & Rollini, 2001). EPSs constitute different classes of
organic macromolecules such as polysaccharides, proteins, nucleic
acids, phospholipids and other polymeric compounds thereby car
rying different organic functional groups such as acetyl, succinyl
or pyruvyl and some inorganic constituent like sulfate (Nielsen &
Jahn, 1999). Depending on their location, EPSs occur in two forms
either in capsular (polymers being closely associated with the cell
surface) or slimy polysaccharides (loosely associated with the cell
surface) (Costerton, 1999).
In recent years, interest in the exploitation of valuable EPSs
has been considerably increased towards polysaccharide produc
ing bacteria and cyanobacteria for various industrial applications
(Mishra & Jha, 2009). The EPSs produced by marine bacteria have
∗ Corresponding author. Tel.: +91 278 256 5801/256 3805x614;
fax: +91 278 256 6970/256 7562.
Email address: [email protected] (C.R.K. Reddy).
been utilized as ingredients in food products, pharmacy, petroleum
industry and emulsification of crude oil, hydrocarbons, vegetable,
mineral oils and bioremediation agents in environment manage
ment system (Costerton, 1999; Jia, Yu, Lin, & Dai, 2007). Apart
from these, they have also been implicated in biofilm formation
in marine ecosystem thereby enhancing the survival of microbes
under abiotic and biotic stress by influencing their physicochemical
environment (Bhaskar & Bhosle, 2005; Duan et al., 2008).
The bacteria are ubiquitous colonizers on the surface of seaweed
and are reported to play important roles in their growth, develop
ment and reproduction (Marshall, Joint, Callow, & Callow, 2006).
Similar findings have been demonstrated with Bacillus licheniformis
which influence the life history and developmental morphology of
Ulva fasciata (Singh, Mantri, Reddy, & Jha, 2011) in corresponding
to increase the zoospore production and in attaining the normal
foliose morphology. Further, seaweed–bacteria interactions are
partly attributed to the extracellular substances secreted by associ
ated marine bacteria. The previous reports reveal that the addition
of supernatants of morphogenesisinducing bacterial isolates were
capable of giving the same effect (Marshall et al., 2006; Tatewaki,
Provasoli, & Pintner, 1983).
In this study, an attempt was made to extract and character
ize the chemical and physical properties of EPS produced by an
endophytic bacterium B. licheniformis associated with Gracilaria
dura with the help of advanced analytical techniques. Despite the
biotechnological potential of these biopolymers from marine and
estuarine environments, the role of EPS in seaweed–bacteria inter
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doi:10.1016/j.carbpol.2010.12.061
Author's personal copy
Carbohydrate Polymers 83 (2011) 891–897
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An alkalihalotolerant cellulase from Bacillus flexus isolated from green seaweed
Ulva lactuca
Nitin Trivedi, Vishal Gupta, Manoj Kumar, Puja Kumari, C.R.K.Reddy ∗, B. Jha
Discipline of Marine Biotechnology and Ecology, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research (CSIR),
Bhavnagar 364021, India
a r t i c l e i n f o
Article history:
Received 10 May 2010
Received in revised form 25 August 2010
Accepted 31 August 2010
Available online 6 September 2010
Keywords:
Cellulose
Cellulase
Haloalkali tolerance
Marine habitat
a b s t r a c t
An extracellular alkalihalotolerant cellulase from the strain Bacillus flexus NT isolated from Ulva lactuca
was purified to homogeneity with a recovery of 25.03% and purity fold of 22.31. The molecular weight of
the enzyme was about 97 kDa and the Vmax and Km was 370.17 U/ml/min and 6.18 mg/ml respectively.
The optimum pH and temperature for enzyme activity was 10 and 45 ◦C respectively. The enzymatic
hydrolysis of the CMC was confirmed with GPC and GCMS analysis. The stabilized activity of the enzyme
even at high pH of 9.0–12.0 and residual activity of about 70% at salt concentration (NaCl 15%) revealed for
its alkalihalotolerance nature. The metal ions Cd2+ and Li1+ were found as inducers while Cr2+, Co2+, Zn2+
and metal chelator EDTA have significantly inhibited the enzyme activity. Enzyme activity was insensitive
to ethanol and isopropanol while partially inhibited by acetone, cyclohexane and benzene.
© 2010 Elsevier Ltd. All rights reserved.
1. Introduction
The industrial and agriculture wastes contain considerable
amounts of cellulose that can effectively be utilized either as a
major source of energy feedstock or as a raw material for production
of high value chemicals (Cherry & Fidantsef, 2003; Kim, Yoo, Oh, &
Kim, 2003). Cellulose, the most abundant carbohydrate in nature,
is a linear polysaccharide of repeating units of glucose linked with
1,4 bacetal bond and mainly forms the primary structural cell wall
component in both the lower and higher plants (Saha, Roy, Sen,
& Ray, 2006). The herbaceous and woody plants are the primary
sources of cellulose intact with complex hemicelluloses, lignin and
pectin. Conversely, seaweeds contain mainly acellulose without
much complex lignin thus differentiating them those of terrestrial
plants and make them preferential source of cellulose (Siddhanta
et al., 2009). The efficient hydrolytic conversion of cellulose into
its monomers, i.e. glucose as source for highenergy molecule will
facilitate to meet the future energy need and also will be an alter
nate to starch. Chemical hydrolysis (acid hydrolysis) is one of the
viable methods currently being employed as a promising means of
producing sugar from cellulose. The combination of high tempera
tures and strong acids in acid hydrolysis leads to the degradation of
products, accumulation of nonsugar byproducts (such as inhibitors
∗ Corresponding author. Tel.: +91 278 256 5801/256 3805x614; fax: +91 278 256
6970/256 7562.
Email address: [email protected] ( C.R.K.Reddy).
to subsequent chemical and biological conversion), and also pose
problem of recovery of reaction agents and resulting saccharides
(Sasaki et al., 1998).
The microorganisms with potential cellulolytic activities could
provide unique opportunity towards the biodegradation of cel
lulosic matter through efficient enzymatic conversion into high
energetic molecules (Wen, Liao, & Chen, 2005). Cellulases are
inducible enzymes synthesized by microorganisms during their
growth on the media containing cellulose as a sole source for
carbon (Lee & Koo, 2001). At commercial scale, cellulases have
been obtained mainly from fungal species of Aspergillus and Tri
choderma due to their high activity but at moderate temperature
(Nandakumar, Thankur, Raghavarao, & Ghildyal, 1994). Several bac
terial genera reported for cellulolytic activities include Bacillus,
Clostridium, Cellomonas, Rumminococcus, Alteromonas, Acetivibrio,
Bacteriodes (Roboson & Chambliss, 1989). Industrial applications
of cellulases have been potentially utilized in leather, textile, agri
culture, food, paper and pulp industries (Bhat, 2000; Kim, Hur, &
Hong, 2005). The industrial utility of cellulase enzymes can fur
ther be improved by investigating the functional efficiency of these
enzymes under extreme conditions of temperature and pH. In
comparison to terrestrial environment, marine habitat with hyper
variable conditions could represent the novel functional abilities of
the microbes that can be further elucidated for their potential as
source of extracellular enzymes.
This study describes the potential of marine bacteria B. flexus
as a source of extracellular cellulase with promising applications in
various industries. The strain was studied for salt tolerance with the
01448617/$ – see front matter © 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.carbpol.2010.08.069
Author's Personal Copy
Technical Note
Solvent tolerant marine bacterium Bacillus aquimaris secreting organic solventstable alkaline cellulase
Nitin Trivedi, Vishal Gupta, Manoj Kumar, Puja Kumari, C.R.K. Reddy ⇑, Bhavanath Jha
Discipline of Marine Biotechnology and Ecology, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research (CSIR), Bhavnagar 364 021, India
a r t i c l e i n f o
Article history:
Received 19 July 2010
Received in revised form 1 February 2011
Accepted 1 February 2011
Keywords:
Solvent tolerance
Bacillus aquimaris
Fatty acid
Cellulase
Ionic liquids
a b s t r a c t
The organic solvent tolerant bacteria with their physiological abilities to decontaminate the organic pol-
lutants have potentials to secrete extracellular enzymes of commercial importance. Of the 19 marine bac-
terial isolates examined for their solvent tolerance at 10 vol.% concentration, one had the significant
tolerance and showed a relative growth yield of 86% for acetone, 71% for methanol, 52% for benzene,
35% for heptane, 24% for toluene and 19% for ethylacetate. The phylogenetic analysis of this strain using
16S rDNA sequence revealed 99% homology with Bacillus aquimaris. The cellulase enzyme secreted by this
strain under normal conditions showed an optimum activity at pH 11 and 45 °C. The enzyme did show
functional stability even at higher pH (12) and temperature (75 °C) with residual activity of 85% and
95% respectively. The enzyme activity in the presence of different additives were in the following order:
Co+2 > Fe+2 > NaOCl2 > CuSO4 > KCl > NaCl. The enzyme stability in the presence of solvents at 20 vol.%
concentration was highest in benzene with 122% followed by methanol (85%), acetone (75%), toluene
(73%) and heptane (42%). The pre-incubation of enzyme in ionic liquids such as 1-ethyl-3-methylimida-
zolium methanesulfonate and 1-ethyl-3-methylimidazolium bromide increased its activity to 150% and
155% respectively. The change in fatty acid profile with different solvents further elucidated the physio-
logical adaptations of the strain to tolerate such extreme conditions.
Ó 2011 Elsevier Ltd. All rights reserved.
1. Introduction
The environment nowadays is increasingly contaminated with
highly toxic monocyclic aromatic hydrocarbons (benzene, toluene,
ethyl benzene and xylenes) as a consequence of rapidly developing
petrochemical and power generating industries worldwide (Wang
et al., 2008). These hydrocarbons are collectively grouped under
BTEX and reported to be carcinogens and pose serious concern to
human health and other kinds of life (Fang et al., 2004). The marine
environment receives about 700 m3 of oil and a variety of hydro-
carbons annually from shipping activities, accidental fuel spills,
and petrochemical based industries (Islam and Tanaka, 2004).
Organic solvents are extremely toxic to cells by virtue of their
ability to disrupt the normal functioning of biological membranes.
The toxicity of solvents is classified on the basis of the measured
values of their partition coefficient in n-octanol and water and
estimated in terms of log Pow value. Solvents with a low log Pow(1.5–4.0) are considered extremely toxic, while others with a high-
er log Pow are less toxic (Inoue et al., 1991). The toxicity level of
organic solvents leads to the cell death by dissolving and disorga-
nizing the cell membrane and finally causes the loss of lipids and
proteins. The microbes dwelling in such toxic organic solvent
habitats can effectively be utilized for bioremediation and various
biotransformation processes of industrial effluents (Inoue et al.,
1991; Ramos et al., 1995; Na et al., 2005; Fang et al., 2006; Chen
et al., 2009). Micro-organisms develop versatile mechanisms such
as rigidification and alteration of cell membrane, or by forming
vesicles to resist the entry of organic solvents (Heipieper et al.,
2007; Segura et al., 2008). The cis–trans isomerization of fatty acids
(FAs) has also been reported as adaptive mechanism in Pseudomo-
nas and Vibrio (Ramos et al., 1997; Heipieper et al., 2003). The other
adaptive mechanisms include modification of lipopolysaccharide
or the porines, active export of the solvent and transformation of
the solvent (Isken and de Bont, 1998). The organic solvent tolerat-
ing microbes particularly the BTEX degraders have been investi-
gated from terrestrial environments that includes largely from
the soil, sewage and also from anaerobic habitat (Botton and
Parsons, 2006). However, only a few micro-organisms from marine
habitat have been investigated for solvent tolerance. Marine
microbes are often exposed to diverse conditions ranging from
extreme temperature, pressure, salinity, pH, photo-radiations, hea-
vy metals, organic solvents and hydrocarbons. In addition to phys-
iological adaptive characteristics, synthesis of exozymes is one of
the notable adaptive catabolic mechanisms of micro-organisms
0045-6535/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.chemosphere.2011.02.006
⇑ Corresponding author. Tel.: +91 278 256 5801/3805x614; fax: +91 278 256
6970/7562.
E-mail address: [email protected] (C.R.K. Reddy).
Chemosphere 83 (2011) 706–712
Contents lists available at ScienceDirect
Chemosphere
journal homepage: www.elsevier .com/locate /chemosphere
Author's personal copy
Growth and agarose characteristics of isomorphic gametophyte (male and female)
and sporophyte of Gracilaria dura and their marker assisted selection
Vishal Gupta, Ravi S. Baghel, Manoj Kumar, Puja Kumari, Vaibhav A. Mantri, C.R.K. Reddy ⁎, Bhavnath Jha ⁎
Discipline of Marine Biotechnology and Ecology, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research (CSIR), Bhavnagar 364021, India
a b s t r a c ta r t i c l e i n f o
Article history:
Received 7 February 2011
Received in revised form 4 June 2011
Accepted 7 June 2011
Available online 15 June 2011
Keywords:
Agarose
Chromosome
Gracilaria dura
Marker assisted selection
Life cycle stage
The characteristics of agarose and growth for three isomorphic life phases of G. durawith their bio-molecular
marker assisted selection have been described in this study. The tetrasporophyte showed superior quality of
agarose over gametophytes and recorded growth rate was highest for females. The genetic relatedness
studied with ISSR markers showed quadratic line of correlation between these phases (R2=1). Their genetic
diversity determinants as percentage of polymorphic loci (PPL), average heterozygosity (He) and Shannon's
Weaver index (I) were 55.55%, 0.5±0.07 and 0.33 respectively. The cytological analysis for chromosome
count revealed 8 chromosomes in haploid gametophytic thallus (N) and 16 for diploid tetrasporophyte (2N)
together with genetic structure analysis confirmed to their sexual mating behaviour. Their marker assisted
selection based on ISSR generated characteristic band of 430 bp specific to male, 860 bp for female and two
bands of 800 and 1600 bp for tetrasporophytic thallus from primer ‘A’. Similarly ISSR primer ‘E’ also generated
bands specific to male, female and tetrasporophytes while others gave bands specific to either of life phase.
Interestingly, endogenic ABA content was significantly higher for haploid gametophytes (female more than
male) than diploid tetrasporophyte while no significant difference was observed in IBA content. Thus the
study described not only the features of three life phases of G. dura but also reliable biomarkers for
differentiating such isomorphic life phases which could be beneficial for the selection of cultivar and in
breeding programmes.
© 2011 Elsevier B.V. All rights reserved.
1. Introduction
Among the red algae, genus Gracilaria constitutes important
agarophytes with more than 150 species reported across the world
(Byrne et al., 2002). The development of improved processing
technologies for agar extraction has increased the annual harvest
rate of this genus. The agar industries throughout the world consume
over 72,300 dry tonnes of agarophytes annually to produce ca.
9600 tonnes of agar worth of US$173 million (Bixler and Porse,
2011). Of this, Gracilaria alone accounts for about 80% of the world's
agar market with a value of US$ 138 million (Bixler and Porse, 2011).
The increasing demand for agar worldwide coupled with short
supplies of raw material from wild stocks has led to the development
of viablemethods for their large scale cultivation in the sea (Peng et al.,
2009). The cultivation of Gracilaria for commercial purposes is being
carried out in several countries including Chile, China and Taiwan and
on pilot scale in Namibia, Venezuela, Mexico and India.
Gracilaria dura (C. Agardh) J. Agardh from the Indian waters has
been reported to produce superior quality agarose (1%
gelN1900 g cm−2) suitable for biotechnological applications (Meena
et al., 2007). Like other tropical seaweeds the limited distribution
coupled with short lifespan prevented its commercial utilisation thus
underlie the need for their cultivation. Further, selection of cultivars
or life phases with superior traits of growth and phycocolloid will
ascertain the economical viability of resources.
The genus Gracilaria has a characteristic Polysiphonia type life cycle
with an alternation of isomorphic gametopyhtic (male and female)
and tetrasporophytic generations (Engel et al., 2004). Moreover,
isomorphic life phases with high resemblances in their morphological
features have frequently misled the identification. Thus, segregation
of life phases at early stages of their development is desired for
cultivar selection, breeding and other biotechnological interventions
aimed at genetic improvement.
In higher plants, several physiological processes are controlled
with synchronised hormonal signals (Stirk et al., 2009). The reported
implication of plant growth regulators (PGRs) seems to be very
promising since some of these compounds can be used as potential
markers for segregating the isomorphic life phases of species such as
G. dura. Along with the PGRs, the adoption of molecular sex-linked
markers offers additional tools for differentiation of life phases of
species even at their juvenile stages dispensing the long wait till the
development of reproductive bodies (Sim et al., 2007).
The RAPD based molecular markers have been extensively
employed for determination of sex in higher plants (Chaves-Bedoya
Aquaculture 318 (2011) 389–396
⁎ Corresponding authors. Tel.: +91 278 256 5801/256 3805x614; fax: +91 278 256
6970/256 7562.
E-mail address: [email protected] (C.R.K. Reddy).
0044-8486/$ – see front matter © 2011 Elsevier B.V. All rights reserved.
doi:10.1016/j.aquaculture.2011.06.009
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Genetic analysis and marker assisted identification of life phases
of red alga Gracilaria corticata (J. Agardh)
Ravi S. Baghel • Puja Kumari • A. J. Bijo •
Vishal Gupta • C. R. K. Reddy • B. Jha
Received: 5 June 2010 / Accepted: 16 November 2010 / Published online: 30 November 2010
Ó Springer Science+Business Media B.V. 2010
Abstract The present study firstly reports the cytological
and molecular marker assisted differentiation of isomor-
phic population of Gracilaria corticata (J. Agardh) with
inter and intra-phasic genetic diversity analysis using ISSR
markers. The genetic diversity of inbreeding population of
G. corticata as determined in terms of percentage of
polymorphic loci (PPL), average heterozygosity (He) and
Shannon’s Weaver index (I) were 59.80, 0.59 and 1.21,
respectively. The inter-phasic pair-wise average polymor-
phism were found to be 31.6% between male and female,
24.0% in male and tetrasporophyte and 25.3% in female
and tetrasporophyte. The intra-phasic average polymor-
phisms were calculated as a maximum of 5.5% between
females, 4.2% between males and the lowest 2.4% between
tetrasporophytes. The primer 10 generated a marker of
800 bp specific to male and 650 bp to female gametophyte,
while the primer 17 generated a marker of 2,500 bp spe-
cific to tetrasporophyte. Both the UPGMA based dendro-
gram and PCA analysis clustered all the three life phases
differentially as distinct identity. Cytological analysis by
chromosome count revealed 24 chromosomes in both
haploid male and female gametophytes (N) and 48 for
diploid (2 N) tetrasporophyte further confirming their
genetic distinctness. The life phase specific markers
reported in this study could be of help in breeding pro-
grammes where differentiation of life phases at the early
developmental stages is crucial.
Keywords Gracilaria corticata Life phase
Inter-Simple Sequence Repeat (ISSR) markers
Genetic structure Chromosome
Introduction
Among the seaweeds, Gracilaria is the third largest genus
with more than 150 species reported across the world and
consists of many commercially important agarophytes [2].
At present, it contributes to more than half of the global
agar industry requirement [18]. Farming of Gracilaria is
being carried out in several countries including the Phil-
ippines, Chile, China, Korea, Indonesia, Namibia, Vietnam
and Argentina at a commercial scale [14] to complement
the growing raw material demand by world agar industry.
Of the Gracilaria species, G. corticata which commonly
inhabit in the intertidal pools and lower intertidal zone has
been reported as a potential source of raw material for food
grade agar [1, 8]. G. corticata from the west coast of India
has been reported to yield food grade agar of about 16%
with the gel strength of 100 g/cm2 [15].
The G. corticata has an isomorphic triphasic life cycle
wherein an alternation of morphologically inseparable yet
genetically distinct generations of haploid and diploid
phases occurs. Of the three life phases only the haploid
female gametophytes can easily be identified by the pres-
ence of cystocarps on the surface of thallus after fertiliza-
tion. The other two life phases i.e. male gametophyte and
tetrasporophyte can be identified microscopically only after
reproductive maturity. At present, there are no reliable
Electronic supplementary material The online version of thisarticle (doi:10.1007/s11033-010-0543-y) contains supplementarymaterial, which is available to authorized users.
R. S. Baghel P. Kumari A. J. Bijo V. Gupta
C. R. K. Reddy (&) B. Jha
Discipline of Marine Biotechnology and Ecology,
Central Salt and Marine Chemicals Research Institute,
Council of Scientific and Industrial Research (CSIR),
Bhavnagar 364021, India
e-mail: [email protected]
123
Mol Biol Rep (2011) 38:4211–4218
DOI 10.1007/s11033-010-0543-y
Original Article
Assessment of nutrient composition and antioxidant potential of Caulerpaceae
seaweeds
Manoj Kumar, Vishal Gupta, Puja Kumari, C.R.K. Reddy *, B. Jha
Discipline of Marine Biotechnology and Ecology, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research (CSIR), Bhavnagar 364021, India
1. Introduction
An increasing human population, global climate change and the
diversificationof terrestrial food resources for energyneeds in recent
times have raised serious global food security concerns (Rosegrant
and Cline, 2003). Further, the globalization of markets has also
brought about an increasing globalization of foods, diminishing the
boundaries of human races andgeographical regions of the countries
throughout the world. There has also been a quest to explore and
utilize foods from non-conventional sources, of both terrestrial and
marine origin, to enhance and supplement the nutritional quality of
human foods. Also, this in turn eases off the growing burden on
traditional foods. Marine macroalgae, commonly known as sea-
weeds, are one of the living renewable resources of the oceans with
potential food applications. Consumption of seaweeds as sea
vegetables in human diets has been the common practice in several
Asian countries (Nisizawa, 2002). Presently, interest in supplement-
ing the human foods with antioxidants particularly from natural
sources has been on the raise as synthetic antioxidants have been
suspected to be a possible cause for liver damage and carcinogenesis
(Farag et al., 2003; Tang et al., 2001). Therefore, there is a need for
isolationandcharacterizationofantioxidantshavingleastsideeffects
from natural sources as an alternative to synthetic antioxidants.
The previous studies have demonstrated the potential of
enzymatic superoxide dismutase (SOD), catalase (CAT), ascorbate
peroxidase (APX) and glutathione reductase (GR) and non-
enzymatic (polyphenols, glutathione, ascorbic acid and carotenoids)
antioxidants scavenging the reactive oxygen species (ROS) thus
relieving from the oxidative stresses and other associated health
risks such as cancer, coronary heart diseases, neurodegenerative
diseases and inflammation (Duan et al., 2006; Kuda and Ikemori,
2009; Nagai and Yukimoto, 2003). The recent findings have also
revealedavailability ofusefulmetaboliteswithmedicinal properties
from some marine biota (Blunt et al., 2005; Mayer et al., 2009).
Recently, the genus Caulerpa has attracted the attention of
researchers due to its important secondary metabolite caulerpe-
nyne (CYN) that is reported to exhibit the antineoplastic,
antibacterial and antiproliferative activities (Barbier et al., 2001;
Cavas et al., 2006). Further, it has also been shown to inhibit the cell
division of sea urchin eggs as well as cancer cell lines (Fischel et al.,
1995; Lemee et al., 1993). Three species of Caulerpa namely C.
racemosa, C. scalpelliformis and C. veravelensis have been found
growing luxuriantly in the intertidal region during October–
February along theVeraval coast of Gujarat (north-western coast of
India). Among these three species, C. racemosa with a wide
Journal of Food Composition and Analysis 24 (2011) 270–278
A R T I C L E I N F O
Article history:
Received 17 July 2009
Received in revised form 5 July 2010
Accepted 31 July 2010
Available online 8 December 2010
Keywords:
Antioxidants
Biochemical constituents
Caulerpa
Minerals
Nutritional supplement
Pigments
Polyunsaturated fatty acids
Seafood
Food analysis
Food composition
A B S T R A C T
The proximate nutrient composition, mineral contents, enzymatic and non-enzymatic antioxidant
potential of three Caulerpa species were investigated. All three species were high in ash (24.20–33.70%)
and carbohydrate content (37.23–48.95%) on dry weight basis (DW). The lipid content ranged between
2.64 and 3.06% DW. The mineral contents varied marginally among the species but were in the order of
Na > K > Ca >Mg. The Na/K ratio among the species varied from 1.80 to 2.55 and was lowest in C.
scalpelliformis. A 10 g DW of Caulerpa powder contains 11–21% Fe, 52–60% Ca and 35–43% Mg, which is
higher than the recommended daily allowance (RDA), compared with non-seafood. The percentage sum
of PUFAs (C18:2, C18:3, C20:4 and C20:5) in total fatty acids was highest in both C. scalpelliformis
(39.25%) and C. veravelensis (36.73%) while it was the lowest in C. racemosa (24.50%). The n 6/n 3 ratio
among the species varied from 1.44 to 7.72 and remained within the prescribed WHO standards (<10).
Further, the higher enzymatic dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and
glutathione reductase (GR) and non-enzymatic antioxidant potential of Caulerpa species found in the
present study confirm their usefulness in terms of nutrients and antioxidants.
ß 2010 Elsevier Inc. All rights reserved.
* Corresponding author. Tel.: +91 278 256 5801/256 3805x614;
fax: +91 278 256 6970/256 7562.
E-mail address: [email protected] (C.R.K. Reddy).
Contents lists available at ScienceDirect
Journal of Food Composition and Analysis
journa l homepage: www.e lsev ier .com/ locate / j fca
0889-1575/$ – see front matter ß 2010 Elsevier Inc. All rights reserved.
doi:10.1016/j.jfca.2010.07.007
Differential responses to cadmium induced oxidative stress
in marine macroalga Ulva lactuca (Ulvales, Chlorophyta)
Manoj Kumar • Puja Kumari • Vishal Gupta •
P. A. Anisha • C. R. K. Reddy • Bhavanath Jha
Received: 7 January 2010 / Accepted: 12 January 2010 / Published online: 30 January 2010
Ó Springer Science+Business Media, LLC. 2010
Abstract This study describes various biochemical
processes involved in the mitigation of cadmium
toxicity in green alga Ulva lactuca. The plants when
exposed to 0.4 mM CdCl2 for 4 days showed twofold
increase in lipoperoxides and H2O2 content that
collectively decreased the growth and photosynthetic
pigments by almost 30% over the control. The
activities of antioxidant enzymes such as superoxide
dismutase (SOD), ascorbate peroxidase (APX), glu-
tathione reductase (GR) and glutathione peroxidase
(GPX) enhanced by twofold to threefold and that of
catalase (CAT) diminished. Further, the isoforms of
these enzymes, namely, Mn-SOD (*85 kDa), GR
(*180 kDa) and GPX (*50 kDa) responded specif-
ically to Cd2? exposure. Moreover, the contents of
reduced glutathione (3.01 fold) and ascorbate
(1.85 fold) also increased substantially. Lipoxyge-
nase (LOX) activity increased by two fold coupled
with the induction of two new isoforms upon Cd2?
exposure. Among the polyunsaturated fatty acids,
although n - 3 PUFAs and n - 6 PUFAs (18:3n - 6
and C18:2n - 6) showed relatively higher contents
than control, the latter ones showed threefold increase
indicating their prominence in controlling the cad-
mium stress. Both free and bound soluble putrescine
increased noticeably without any change in spermi-
dine. In contrast, spermine content reduced to half over
control. Among the macronutrients analysed in
exposed thalli, the decreased K content was accom-
panied by higher Na and Mn with no appreciable
change in Ca, Mg, Fe and Zn. Induction of antioxidant
enzymes and LOX isoforms together with storage of
putrescine and n - 6 PUFAs in cadmium exposed
thallus in the present study reveal their potential role in
Cd2? induced oxidative stress in U. lactuca.
Keywords Antioxidant enzymes
Cadmium LOX Minerals Oxidative stress
PUFAs Ulva lactuca
Introduction
Of the toxic substances contaminating the aquatic
environment, heavy metals particularly cadmium,
lead and mercury are of great concern for humans as
well as for the environment because of their acute
toxicity and high mobility in food chain (Sokolova
et al. 2005). Cadmium (Cd2?), with no reported
biological function except one occasion as a cofactor
for carbonic anhydrase in marine diatom (Lane and
M. Kumar P. Kumari V. Gupta
C. R. K. Reddy (&) B. Jha
Discipline of Marine Biotechnology and Ecology, Central
Salt and Marine Chemicals Research Institute, Council
of Scientific and Industrial Research (CSIR), Bhavnagar
364021, India
e-mail: [email protected]
P. A. Anisha
School of Environmental Studies, Cochin University
of Science and Technology, Cochin, India
123
Biometals (2010) 23:315–325
DOI 10.1007/s10534-010-9290-8
Author's personal copy
Biochemical responses of red alga Gracilaria corticata (Gracilariales, Rhodophyta) to
salinity induced oxidative stress
Manoj Kumar, Puja Kumari, Vishal Gupta, C.R.K. Reddy ⁎, Bhavanath Jha
Discipline of Marine Biotechnology and Ecology, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research (CSIR), Bhavnagar 364021, India
a b s t r a c ta r t i c l e i n f o
Article history:
Received 26 February 2010
Received in revised form 31 May 2010
Accepted 1 June 2010
Keywords:
Antioxidant enzymes
Gracilaria corticata
Minerals
Oxidative stress
Phycobiliproteins
PUFAs
Salinity stress
The biochemical responses of Gracilaria corticata (J. Agardh) J. Agardh to salinity induced oxidative stress were
studied following the exposure to different salinities ranging from 15, 25, 35 (control), 45 to 55 in laboratory
conditions. The growth was highest under 25 (3.14±0.69% DGR) and 35 (3.58±0.32% DGR) and decreased
significantly in both extreme lower (15) and hyper (55) salinities. Both phycoerythrin (PE) and
allophycocyanin (APC) were significantly higher in hyper-salinity (45) with an increase of almost 70% and
52% from their initial contents. Conversely, the level of increase of the same in hypo-salinitieswas considerably
lower as compared with that of hyper-salinity. Both hypo- and hyper-salinity treatments induced almost two
fold increase in the contents of polyphenols, proline and the activities of antioxidative enzymes such as
superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR) especially for 6 d
exposure. The Na+ ions readily displaced the K+ and Ca2+ from their uptake sites at extreme hyper-salinity
(55) and accounted for substantial increase in the ratio of Na+/K+ and Na+/Ca2+ that impeded the
growth under long term exposure (N6 d). The survivability at salinity 45 evenwith considerably higher ratio of
Na+/K+ and Na+/Ca2+ suggests the compartmentalization of Na+ into the vacuoles. Further, the micro
nutrients such as Zn, Fe and Mn were decreased at both high and low end salinities with highest at extreme
hyper-salinity. The C18:1(n−9) cis, C18:2(n−6), C18:3(n−3) and C20:4(n−6) were found in significant
amounts in hyper-salinities. The C18:1(n−9) cis in particular increased by 60.25% and 70.51% for salinities 45
and 55, respectively from their initial amounts. The ratio of total unsaturated to saturated fatty acids (UFA/SFA)
also increased linearly with increasing salinity. These results collectively suggest the potential role of
antioxidative enzymes, phycobiliproteins, PUFAs and mineral nutrients to combat the salinity induced
oxidative stress in G. corticata.
© 2010 Elsevier B.V. All rights reserved.
1. Introduction
The red alga Gracilaria corticata (J. Agardh) J. Agardh is one of the
common algae of the Indian coast and occurs predominantly in the
lower littoral zone. It also inhabits occasionally in the intertidal rock
pools as submerged population. The intertidal algae often get exposed
to the atmosphere periodically during low tide regimes and experi-
ence an oxidative stress on regular basis with the turning tides. In
marine waters, salinity around 35 is the most common, but it could
also vary from 10 to 70 as a result of evaporation or precipitation/
freshwater influxes (Graham and Wilcox, 2000). Osmotic stress most
often resulting from fluctuating salinities exerts considerable oxida-
tive stress on seaweeds in the intertidal zone. The previous studies
have investigated the responses of estuarine macroalgae for either
individual or combined abiotic factors (light, pH, temperature,
nutrient load and salinity) in the context of growth andphotosynthetic
performance (Macler, 1988; Dawes et al., 1999; Israel et al., 1999; Choi
et al., 2006; Phoopronget al., 2007). Subsequent studies have also dealt
with the possible effects of environmental stresses on floristic
variations of intertidal benthic macro algal communities (Helmuth
et al., 2005).
It has been suggested that instant responses of marine plants to
adverse environmental conditions involve excess production of
reactive oxygen species (ROS) such as hydrogen peroxide (H2O2),
singlet oxygen (1O2), superoxide radical (O2•−) and hydroxyl radical
(OH−) (Dring, 2006). Increased physiological stress conditions lead to
the rapid formation of ROS that reacts with most cellular components
and thus they need to be neutralized instantly once formed.
Acclimation to altered osmotic conditions particularly to salinity
induced stress involves changes in physiological processes including
antioxidant enzymes [superoxide dismutase (SOD), catalase (CAT),
ascorbate peroxidase (APX) and glutathione reductase (GR)] and non-
enzymatic antioxidants (ascorbate, glutathione and carotenoids).
All these processes function in coordinated manner in order to
alleviate the cellular hypo/hyper osmolarity, ion disequilibrium and
Journal of Experimental Marine Biology and Ecology 391 (2010) 27–34
⁎ Corresponding author. Tel.: +91 278 256 5801/256 3805x614; fax: +91 278 256
6970 / 256 7562.
E-mail address: [email protected] (C.R.K. Reddy).
0022-0981/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.jembe.2010.06.001
Contents lists available at ScienceDirect
Journal of Experimental Marine Biology and Ecology
j ourna l homepage: www.e lsev ie r.com/ locate / jembe
Purification and partial characterization of an extracellular
alginate lyase from Aspergillus oryzae isolated
from brown seaweed
Ravindra Pal Singh & Vishal Gupta & Puja Kumari &
Manoj Kumar & C. R. K. Reddy & Kamalesh Prasad &
Bhavanath Jha
Received: 15 April 2010 /Revised and accepted: 9 August 2010 /Published online: 2 September 2010# Springer Science+Business Media B.V. 2010
Abstract The extracellular enzyme alginate lyase produced
from marine fungus Aspergillus oryzae isolated from brown
algaDictyota dichotoma was purified, partially characterized,
and evaluated for its sodium alginate depolymerization
abilities. The enzyme characterization studies have revealed
that alginate lyase consisted of two polypeptides with about
45 and 50 kDa each on 10% sodium dodecyl sulfate
polyacrylamide gel electrophoresis and showed 140-fold
higher activity than crude enzyme under optimized pH (6.5)
and temperature (35°C) conditions. Zn2+, Mn2+, Cu2+, Mg2+,
Co2+ and NaCl were found to enhance the enzyme activity
while (Ca2+, Cd2+, Fe2+, Hg2+, Sr2+, Ni2+), glutathione, and
metal chelators (ethylenediaminetetraacetic acid and eth-
ylene glycol tetraacetic acid) suppressed the activity.
Fourier transform infrared and thin-layer chromatography
analysis of depolymerized sodium alginate indicated the
enzyme specificity for cleaving at the β-1,4 glycosidic
bond between polyM and polyG blocks of sodium alginate
and therefore resulted in estimation of relatively higher
polyM content than polyG. Comparison of chemical shifts
in 13C nuclear magnetic resonance spectra of both polyM
and polyG from that of sodium alginate also showed
further evidence for enzymatic depolymerization of sodi-
um alginate.
Keywords Alginate lyase . Aspergillus oryzae . Fungus .
PolyM . PolyG . Sodium alginate
Introduction
Alginate occurs as a structural cell wall polysaccharide in a
wide variety of brown seaweeds. It can also be obtained
from bacteria such as Azotobacter vinelandii (Gorin and
Spencer 1966) and Pseudomonas aeruginosa (Evans and
Linker 1973) but with poor gelling characteristics. Alginates
are linear unbranched polymers consisting of 1,4-linked β-D-
mannuronic acid (M) and α-L-guluronic acid (G) blocks,
arranged as either homopolymeric (M–M or G–G blocks) or
heteropolymeric (M–G and G–M blocks) random sequences
(Gacesa 1992). Alginates are commercially important cell
wall polysaccharides and widely used as stabilizers, visco-
sifiers, and gelling agents in diverse products such as food,
beverages, and pharmaceuticals industries (Wong et al.
2000). The polyM has been well studied and reported as a
potent inducer of cytokines under acute inflammatory
responses (Jahr et al. 1997). In contrast, polyG inhibits the
secretion of cytokines, resulting to the alleviation of the
immunostimulation during tissue grafting and other autoim-
mune disorders (Otterlei et al. 1992). The alginate derivatives
with sulfate groups have been reported to have high tumor
inhibition activity against solid sarcoma 180 in vivo (Hu et
al. 2004) in addition to tissue engineering applications
(Kataoka et al. 2004). Further, the depolymerized products
of alginate have also been stated to promote germination,
growth, and development in crop plants (Cao et al. 2007).
Alginate lyases, characterized as either mannuronate (EC
4.2.2.3) or guluronate lyases (EC 4.2.2.11), catalyze the
degradation of alginate. Alginates can be depolymerized
into respective oligosaccharide fragments using either
enzymatic lyases or acid hydrolysis. Alginate lyase uses a
β-elimination in which a non-reducing unsaturated bond is
produced during cleavage of the uronic acid, giving rise to
R. P. Singh : V. Gupta : P. Kumari : M. Kumar :
C. R. K. Reddy (*) : K. Prasad : B. Jha
Discipline of Marine Biotechnology and Ecology, Central Salt and
Marine Chemicals Research Institute, Council of Scientific and
Industrial Research (CSIR),
Bhavnagar 364021, India
e-mail: [email protected]
J Appl Phycol (2011) 23:755–762
DOI 10.1007/s10811-010-9576-9