Variation in chemical componenets of aquacultured Ulva( Chlorophyta )

131 Pages·2014·2.85 MB·English

Variation in chemical components of
aquacultured Ulva (Chlorophyta) in

response to environmental varniables
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Etwarysing Lekraej
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Supervisors: Prof. John J Boltofn (University of Cape Town)
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Assoc Prof. Denzil Beukes
(University of the Western Cape)
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Minor Dissertation presented in partial fulfillment of the requirements for
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the degreei of Master of Science (Applied Marine Science)
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Department of Biological Sciences
Faculty of Science
University of Cape Town

September 2014
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The copyright of this thesis vests in the author. No
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quotation from it or information derived from it is to be

published without full acknowledgeement of the source.
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The thesis is to be used for private study or non-
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commercial research purposes only.

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Published by the Universit y of Cape Town (UCT) in terms
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of the non-exclusive license granted to UCT by the author.
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Plagiarism declaration
I know the meaning of plagiarism and declare that all of the work in the dissertation,
save for that which is properly acknowledged, is my own. Aside from guidance from
my supervisors, I have received no assistance, except as acknowledged.
SIGNATURE: Signature Removed
DATE: 19th September 2014
Table of Contents
List of Tables I
List of Figures I
Acknowledgements VII
Abstract VIII

Chapter 1: Literature Review 1
1.1 The genus Ulva 1
1.2 Distribution 2
1.3 Life History of Ulva 3
1.4 Environmental factors affecting Ulva 5
1.4.1 Light 5
1.4.2 Temperature 7
1.4.3 Salinity 8
1.4.4 Nutrients 9
1.4.5 Effects of heavy metals 11
1.5 Volatile organic compounds 12
1.6 Nutritional content 15
1.7 Ulva in aquaculture 17
1.8 South African abalone industry 20

Chapter 2 22
2.1 Introduction 22
2.2 Materials and Methods 25
2.2.1 Plant material 25
2.2.2 Culture experiment 26
2.2.3 Grazing experiment 27
2.2.3.1 Natural grazing 27
2.2.3.2 Artificial grazing 27
2.2.4 Sample preparation and NMR analysis 28
2.2.5 Data analysis 29
2.3 Results 31
2.3.1 Effects of grazing on Ulva metabolites 31
2.3.2 Effects of nutrient and temperature variations on Ulva metabolites 33
2.3.3 Effects of salinity and temperature variations on Ulva metabolites 34
2.3.4 Principal Components Analysis 36
2.3.4.1 Grazing experiment 37
2.3.4.2 Nutrient experiment 38
2.3.4.3 Salinity experiment 39

2.4 Discussion and Conclusion 42
Recommendations 49
References 50
Appendices 85

I . Whole 1H NMR spectra 85
- Grazing experiment 85
- Nutrient experiment 87
- Salinity experiment 91
II. Normalized 1H NMR spectra 101
- Grazing experiment 101
- Nutrient experiment 103
- Salinity experiment 107
List of tables
Page
Table 1 : Average aldehyde intensity for peak signal at δ 9.76 for the 31
H
nutrient experiment at 10 °C and 15 °C.

Table 2 : Average aldehyde intensity for peak signal at δ 9.76 for the 34
H
salinity experiment at 10 °C and 15 °C.

List of figures
Page
Figure 1 : Typical sporic meiosis life history of Ulva (A) Male and 4
female gametophyte; (B) Biflagellate gamete; (C)
Sporophyte; (D) Quadriflagellate zoospore; 1N- haploid and
2N- diploid. (Source: Beach et al., 1995).

Figure 2 : The cleavage pathway producing DMS and acrylic acid from 14
DMSP by DMSP lyase in seaweed (Source: Van Alstyne et
al., 2001).

Figure 3 : NMR spectrum of Ulva armoricana with an aldehyde peak at 29
9.76 ppm and the DMF peak at 8.03 ppm.

Figure 4 : 1H NMR spectra of Ulva armoricana for grazing experiments 32
with replicate B (maroon) representing natural grazing
experiment and replicate B (green) representing the artificial
grazing experiment from δ -0.20 to δ 12.20 for the whole
H H
spectra and δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure 5 : 1H NMR spectra of Ulva armoricana for low nutrient supply 33
(0 % ES) experiment with replicate B (green) representing
treatment at 10 °C and replicate A (maroon) for treatment at
15 °C from δ -0.20 to δ 12.20 for the whole spectra and δ
H H H
9.00 to δ 10.50 for the aldehyde region.
H

Figure 6 : 1H NMR spectra of Ulva armoricana for high low nutrient 34
supply (100% ES) experiment with replicate A (maroon)
representing treatment at 10 °C and replicate C (blue) for
treatment at 15 °C from δ -0.20 to δ 12.20 for the whole
H H
spectra and δ 9.00 to δ 10.50 for the aldehyde region.
H H

I
Figure 7 : 1H NMR spectra of Ulva armoricana for salinity 25 ‰ 35
treatment with replicate C (blue) representing treatment at 10
°C and replicate B (green) for treatment at 15 °C from δ -
H
0.20 to δ 12.20 for the whole spectra and δ 9.00 to δ 10.50
H H H
for the aldehyde region.

Figure 8 : PCAs of normalized bins values of Ulva armoricana for the 37
grazing experiments with PCA (A; left) representing the
whole spectrum between δ -0.20 to δ 12.20 and PCA (B;
H H
right) representing the aldehyde region between δ 9.00 to δ
H H
10.50.

Figure 9 : PCAs of normalized bins values of Ulva armoricana for the 39
nutrient experiments with PCA (A; left) representing the
whole spectrum between δ -0.20 to δ 12.20 and PCA (B;
H H
right) representing the aldehyde region between δ 9.00 to δ
H H
10.50.

Figure 10 : PCAs of normalized bins values of Ulva armoricana for the 40
salinity experiments at 10 °C with PCA (A; left) representing
the whole spectrum between δ -0.20 to δ 12.20 and PCA
H H
(B; right) representing the aldehyde region between δ 9.00
H
to δ 10.50.
H

Figure 11 : PCAs of normalized bins values of Ulva armoricana for the 41
salinity experiments at 15 °C with PCA (C; left) representing
the whole spectrum between δ -0.20 to δ 12.20 and PCA
H H
(D; right) representing the aldehyde region between δ 9.00
H
to δ 10.50.
H

Figure A1 : 1H NMR spectra of Ulva armoricana for natural grazing 85
experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 2 : 1H NMR spectra of Ulva armoricana for artificial grazing 86
experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 3 : 1H NMR spectra of Ulva armoricana for low nutrient supply 87
(0% ES) at 10 °C experiment with replicates: A (maroon), B
(green) and C (blue) from δ -0.20 to δ 12.20 for the whole
H H
spectra and from δ 9.00 to δ 10.50 for the aldehyde region.
H H

II
Figure A 4 : 1H NMR spectra of Ulva armoricana for high nutrient supply 88
(100% ES) at 10 °C experiment with replicates: A (maroon),
B (green) and C (blue) from δ -0.20 to δ 12.20 for the
H H
whole spectra and from δ 9.00 to δ 10.50 for the aldehyde
H H
region.

Figure A 5 : 1H NMR spectra of Ulva armoricana for low nutrient supply 89
(0% ES) at 15 °C experiment with replicates: A (maroon), B
(green) and C (blue) from δ -0.20 to δ 12.20 for the whole
H H
spectra and from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 6 : 1H NMR spectra of Ulva armoricana for high nutrient supply 90
(100% ES) at 15 °C experiment with replicates: A (maroon),
B (green) and C (blue) from δ -0.20 to δ 12.20 for the
H H
whole spectra and from δ 9.00 to δ 10.50 for the aldehyde
H H
region.

Figure A 7 : 1H NMR spectra of Ulva armoricana for salinity 5 ‰ at 10 91
°C experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 8 : 1H NMR spectra of Ulva armoricana for salinity 10 ‰ at 10 92
°C experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 9 : 1H NMR spectra of Ulva armoricana for salinity 20 ‰ at 10 93
°C experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 10 : 1H NMR spectra of Ulva armoricana for salinity 25 ‰ at 10 94
°C experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 11 : 1H NMR spectra of Ulva armoricana for salinity 35 ‰ at 10 95
°C experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 12 : 1H NMR spectra of Ulva armoricana for salinity 5 ‰ at 15 96
°C experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H
III
Figure A 13 : 1H NMR spectra of Ulva armoricana for salinity 10 ‰ at 15 97
°C experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 14 : 1H NMR spectra of Ulva armoricana for salinity 20 ‰ at 15 98
°C experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 15 : 1H NMR spectra of Ulva armoricana for salinity 25 ‰ at 15 99
°C experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 16 : 1H NMR spectra of Ulva armoricana for salinity 35 ‰ at 15 100
°C experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 17 : Normalized 1H NMR spectra of natural grazing experiments 101
with replicate A (maroon), B (green) and C (blue) from δ -
H
0.20 to δ 12.20 for the whole spectra and δ 9.00 to δ 10.50
H H H
for the aldehyde region.

Figure A 18 : Normalized 1H NMR spectra of artificial grazing experiment 102
with replicate A (maroon), B (green) and C (blue) from δ -
H
0.20 to δ 12.20 for the whole spectra and δ 9.00 to δ 10.50
H H H
for the aldehyde region

Figure A 19 : Normalized 1H NMR spectra of low nutrient supply (0% ES) 103
at 10 °C experiment with replicates: A (maroon), B (green)
and C (blue) from δ -0.20 to δ 12.20 for the whole spectra
H H
and from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 20 : Normalized 1H NMR spectra of high nutrient supply (100% 104
ES) at 10 °C experiment with replicates: A (maroon), B
(green) and C (blue) from δ -0.20 to δ 12.20 for the whole
H H
spectra and from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 21 : Normalized 1H NMR spectra of the low nutrient supply (0% 105
ES) at 15 °C experiment with replicates: A (maroon), B
(green) and C (blue) from δ -0.20 to δ 12.20 for the whole
H H
spectra and from δ 9.00 to δ 10.50 for the aldehyde region.
H H

IV
Figure A 22 : Normalized 1H NMR spectra of the high nutrient supply 106
(100% ES) at 15 °C experiment with replicates: A (maroon),
B (green) and C (blue) from δ -0.20 to δ 12.20 for the
H H
whole spectra and from δ 9.00 to δ 10.50 for the aldehyde
H H
region.

Figure A 23 : Normalized 1H NMR spectra of the salinity 5 ‰ at 10 °C 107
experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 24 : Normalized 1H NMR spectra of the salinity 10 ‰ at 10 °C 108
experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 25 : Normalized 1H NMR spectra of the salinity 20 ‰ at 10 °C 109
experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to δ 12.20 for the whole spectra and
H H
from δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 26 : Normalized 1H NMR spectra of the salinity 25 ‰ at 10 °C 110
experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to 12.20 for the whole spectra and from
H
δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 27 : Normalized 1H NMR spectra of the salinity 35 ‰ at 10 °C 111
experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to 12.20 for the whole spectra and from
H
δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 28 : Normalized 1H NMR spectra of the salinity 5 ‰ at 15 °C 112
experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to 12.20 for the whole spectra and from
H
δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 29 : Normalized 1H NMR spectra of the salinity 10 ‰ at 15 °C 113
experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to 12.20 for the whole spectra and from
H
δ 9.00 to δ 10.50 for the aldehyde region.
H H

Figure A 30 : Normalized 1H NMR spectra of the salinity 20 ‰ at 15 °C 114
experiment with replicates: A (maroon), B (green) and C
(blue) from δ -0.20 to 12.20 for the whole spectra and from
H
δ 9.00 to δ 10.50 for the aldehyde region.
H H

V