The effect of soil extracts on the physiology of

Lyngbya majuscula (Cyanophyta)

Simon Albert

Thesis submitted to the Department of Botany, University of Queensland

as partial fulfilment of BScApp (Env) (Hons)

Supervisors: Dr. Judith O'Neil

A/Prof. William Dennison
Dr. Phil Moody

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Abstract
Recent outbreaks of the cyanobacterium Lyngbya majuscula in Moreton Bay,
Queensland, Australia, have been the impetus for a number of studies to determine
potential triggers for the blooms. The current study assesses the potential for runoff from
various land uses to stimulate the growth of L. majuscula. Soil extracts from eight
representative land uses (including Melaleuca forest, mangroves, pine plantation and
canal development) within the Pumicestone catchment were produced to simulate runoff
events. A biological assay technique was used to determine the potential of each extract
to stimulate L. majuscula productivity. A 102% increase in photosynthetic rate was
recorded (using PAM fluorescence) in response to the cleared pine forest soils. A
significant response was also observed from the forested pine forest soils, with a 33%
increase in photosynthetic rate.

Concomitant increases in the concentration of the

photosynthetic pigment phycoerythrin, may explain this result.

These results were

consistent across a series of experiments, with L. majuscula collected from two different
sites (Eastern Banks and Deception Bay), both showing significant stimulation in
response to the cleared pine soil extract. Phosphorous concentrations in the pine soil
extracts were an order of magnitude higher than the other soil extracts measured, and
were also acidic (pH 3.5-4). Based on spectral and chemical analyses, the forested (Pine
and Melaleuca) soils have high organic carbon and soluble iron content. Parallel studies
indicate these organic rich soil extracts are able to strongly complex iron, providing a
potential transport mechanism for bioavailable iron from the land to reach the L.
majuscula, in the marine environment.

The link made by the current study between

catchment derived compounds and blooms of L. majuscula in Moreton Bay, provides a

focus for potential management actions.

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Introduction
Lyngbya majuscula is a toxic, filamentous marine cyanobacteria within the Oscillatoriacea,

previously cited in literature as Microcoleus lyngbyaceus (Ktzing) (Diaz et al. 1990,

Speziale & Dyck 1992). L. majuscula grows on solid substrates or epiphytically on
seagrass in the coastal zones of many sub-tropical and tropical oceans. Since the early
1990s nuisance blooms of this toxic cyanobacteria have been occurring seasonally in
Moreton Bay, Queensland, Australia (Dennison et al. 1997). While present in small
quantities in many locations throughout Moreton Bay, the L. majuscula bloom events
over the past five years have been both temporally and spatially specific, with Deception
Bay (northern section of Moreton Bay) and the Eastern Banks being the two major bloom
regions.
Lyngbya majuscula blooms typically begin in Moreton Bay in the summer
(December/January) and expand rapidly over the following few months to an area up to
10 km2 (Dennison et al. 1997). This is often followed by a rapid population collapse,
possibly aided by viruses (Hewson et al. 2001). During this cycle it has been observed
that the L. majuscula begins growth from the sediment below the seagrass canopy. As
the 'bloom' develops this benthic mat is able to grow sufficiently to overtop the seagrass
species, with a blanketing effect that can turn anoxic. After continuous periods of high
light, warm temperatures and calm weather the photosynthetic rates of the L. majuscula
mats are sufficiently high to form large amounts of buoyant bubbles within the L.
majuscula matrix. The benthic mats eventually float to the surface of the water and float
freely. This stage may provide a dispersal mechanism for the L. majusucla, enabling it to
spread into other regions of the bay.
Seagrass loss and altered marine plant community structure have been the most
significant initial impacts of Lyngbya majuscula bloom events (Watkinson 2000) to date,
while impacts on biota (particularly turtle and dugong) remain unclear. In addition to
these environmental effects, L. majuscula can have human health impacts. The suite of
toxins within L. majuscula can cause severe skin and eye irritation as well as asthma like
symptoms (Osborne et al. 2001). The potential for serious deleterious environmental and

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human impacts from L. majuscula in Moreton Bay has provided the impetus for a series
of studies to determine potential limiting factors for these blooms.
In general, marine plants within Moreton Bay are nitrogen limited (O'Donohue &
Dennison 1997, Udy & Dennison 1997). Nitrogen limited systems often favour
prokaryotic nitrogen fixers such as cyanobacteria. In the absence of nitrogen limitation,
iron becomes a major limiting factor of biological growth in oceanic systems (Martin &
Gordon 1988, Martin et al. 1990). More recently this has also been found to be the case
in coastal and estuarine ecosystems (Hutchins & Bruland 1998, Hutchins et al. 1998). In
most aquatic environments, cyanobacteria have a high demand for iron (Paerl et al. 1994,
Trick et al. 1995) and phosphorus (Paerl et al. 1987, Sanudo-Wilhelmy et al 2001) for
both photosynthesis and nitrogen fixation. Elevated iron concentrations in laboratory
studies have: increased productivity and phycocyanin production in Oscillatoria tenius
(Trick et al. 1995), elevated nitrogen fixation in Trichodesmium sp. (Rueter et al. 1990)
and increased toxin production by Microcystis aeruginosa (Utkilen & Gjolme 1995).

Iron is one of the most abundant elements on earth, however its insolubility at seawater
pH prevents its availability to marine organisms in most instances (Anderson and Morel
1982). At seawater pH, ferric iron (Fe(III)) is the thermodynamically preferred form,
with free Fe(II) undergoing rapid oxidation and subsequent precipitation of the ferric
form (Byrne & Kester 1976). Cyanobacteria are unable to take up and utilize these
oxides of iron. This oxidation is generally reduced in the presence of organic ligands that
form a complex with the soluble iron making it more persistent in seawater (Emmenegger
et al. 1998, Santana-Casiano et al. 2000). Following reductive processes (e.g. photo-

reduction) to break this complex (Waite & Morel 1984, Wells & Mayer 1991, Voelker et
al. 1997), phytoplankton and cyanobacteria are able to take up soluble iron directly from
the water column (Anderson & Morel 1982). Therefore, the level of bioavailable iron in
seawater can fluctuate greatly depending on the presence of natural complexation agents
such as organics.

Dissolved organic carbon is a natural feature of Australian coastal waters (Kirk 1994),
particularly those areas surrounded by natural vegetation. The leaves of Melaleuca,
Acacia and other lowland vegetation, contain a high proportion of organic carbon (Zoete
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2001). These organics leach from decomposing leaves during rainfall and are either

recycled within the soil system or may be mobilised into the surrounding creeks (Kirk
1994). The organic acids contained within organic-rich terrestrial run-off have been
shown to have strong iron complexation properties (Kuma et al.1996, Matsunaga et al.
1999, Rose et al. unpub. data). Plumes of darkly stained, organic-rich waters have been
observed entering regions surrounding L. majuscula bloom areas in Deception Bay,
particularly following rainfall events and the clearfelling of exotic plantation pine.
Parallel studies investigating the role these organic-rich compounds have as a transport
mechanism for bio-available iron to reach the bloom sites from terrestrial sources, are
currently under way

Coastal waters such as rivers, dams, estuaries are often subject to algal blooms as a result
of nutrient input from terrestrial sources (Mallin et al. 1993). Rivers running through
urbanised catchments generally provide high nutrient loadings as a result of the complex
array of inputs. Forested, undisturbed catchments, generally yield lower water volumes
and more stable, low fluxes of nutrients, reflecting the inherently low nutrient status of
Australian soils (Wild 1958, Beadle 1962, Cambell 1975). Logging of these forests can
greatly alter both water quality and quantity in the surrounding estuaries (Cambell &
Doeg 1989, Rask et al. 1998, Bubb 2000, Roberts 2000), triggering ecological changes
such as algal blooms.

The current study aims to assess the potential for various land uses within the
Pumicestone catchment to be the source of substances stimulating benthic blooms of
Lyngbya majuscula in Deception Bay. Although there are many interactive factors,
identifying identifying the potential causal processes of the bloom, in terms of land use
may help explain recent increases in this noxious cyanobacteria. Thus, rather than purely
scientific knowledge of the nutritional requirement of L. majuscula, this study helps to
provide a more applied and currently much needed result; management focus.

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Study Area
The study was conducted in Moreton Bay, Queensland at two locations, Deception Bay
(27 05 S, 153 09 E) and Eastern Banks (27 26S, 153 24E ) (Figure 1). The
primary focus was in Deception bay and the surrounding catchment (Figure 2). The
coastal system in which Lyngbya majuscula is blooming in Deception Bay is primarily
influenced by a large estuarine passage (Pumicestone Passage) dividing Bribie Island to
the east and the mainland to the west. This passage is in turn fed by 8-10 creeks and
numerous smaller tributaries draining both mainland and island regions. Much of this
area is low-lying and, hence, the catchment boundary is large.
The Pumicestone catchment contains a diversity of horticultural, residential and natural
land uses. The dominant land use within the Pumicestone catchment is exotic pine
plantations (Pinus elliottii) (39%). Other horticultural activities in the Pumicestone
catchment include; sugar cane, strawberries and the remnants of the largest citrus orchard
in the southern hemisphere (Roy family 1960s). Residential areas are primarily
restricted to the dense canal estates on the southern section of Bribie Island and the
adjacent Sandstone Point on the mainland.

Deception
Bay

Moreton
Bay
Eastern
Banks

Figure 1: Satellite imagery depicting sampling regions in relation to Australian and Queensland coastlines.

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10

Bribie
Island
9
1

2
8

Pumicestone
Passage

4 3

7
6

Bloom Site
N

Figure 2: Satellite imagery depicting sampling sites in the Pumicestone catchment.

# Site

Site Description

1 Cleared Pine

Sandy podosol, Pinus elliottii plantation cleared ca. 6 months prior to sampling

2 Intact Pine

Sandy podosol sampled from mature stand of Pinus elliottii

3 Melaleuca

Sandy podosol sampled from mixed forest of Melaleuca and Acacia species

4 Mangrove

Mangrove sediment sampled within an Avicennia marina community

5 Shirley Creek

Sediment sampled from this visibly iron rich creek

6 Sandstone Point

Intertidal marine Sands overlying Landsborough sandstone bedrock

7 Canal Development

Sandy hydrosol sampled from previously dug canal development

8 Coffee Rock

Exposed coffee rock layer in intertidal area of Pumicestone Passage

9 Mellum Creek

Creek draining mainland regions of pine plantation

10 Pine drain

Small earth drain between pine plantation plots on Bribie Is.

Table 1: Description of sampling sites within the Pumicestone catchment region.

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Methods
A bioassay technique was developed using soil extracts from eight representative land
uses (Melaleuca forest, mangroves, cleared and intact pine plantation, canal development,
iron rich creek, marine sediment and coffee rock) within the Pumicestone catchment
(Table 1). These soil extracts were produced to simulate the compounds that these soils
would yield during natural runoff events. Lyngbya majuscula was incubated in dilutions
of these extracts to determine the physiological effects. Secondly, different dilutions of
selected extracts will be used to determine if a concentration threshold exists for L.
majuscula stimulation. Thirdly, L. majuscula from different regions of Moreton Bay was
incubated with selected extracts to determine if the two populations respond similarly.

Soil extract production

Various methods exist for deriving soil extracts. Commonly they are derived through
wetting of intact soil cores until field capacity is reached, to simulate the waterlogging
effect of heavy rainfall. Intact cores are then leached using rainwater for varying periods
of time (up to 40 weeks in some studies) (Khomutova et al. 2000). Although this study
has modified this protocol, comparative studies using leached cores at these sites have
yielded chemically similar extracts (Moody unpub. data).
At sites 1-8, three 400 mm x 90 mm diameter cores of the soil were taken using
polycarbonate corers driven into the ground (Figure 3). The three cores were combined
to reduce spatial variation and the soil structure homogenised. Three kilograms of each
soil type was then mixed with three litres of rainwater and stirred vigorously to produce a
muddy consistency. The mixture was then left in a dark aerobic environment for 24
hours to simulate the water-logging of soil following rainfall. The supernatant was then
decanted off and vacuum filtered through 10 m polycarbonate filters to produce a soil
solution to simulate naturally occurring run-off (Figure 3).

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Bioassays
Bioassays using Lyngbya majuscula were carried out to investigate what effect extracts
from these different land uses had on physiological parameters.

Figure 3: Conceptualisation of methodology used to create soil extracts and assess their effects upon
Lyngbya majuscula physiology.

Bioassay of eight primary land uses

Three replicate one-litre glass beakers were filled with 900 ml seawater from the
Deception Bay bloom area and 100 ml (1:9) of the eight soil solutions, 100 mls of
rainwater was added to the seawater control to ensure consistent salinity (3x9 = 27
beakers). This dilution was based on previous salinity monitoring of the bloom region
indicating a 10% decrease in salinity prior to bloom initiation (Watkinson 2000). Five
cm3 of Lyngbya majuscula was incubated in these solutions beakers over two days under
50% shadecloth. Various physical and biological parameters were assessed over this
incubation period to assess how the extracts affected L. majuscula physiology.
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Bioassay of various extract dilutions

The basic protocol from experiment 1 was replicated. Based on results from the first
experiment, cleared pine, sandstone point and coffee rock extracts were focussed on for
further analysis in this second experiment. Rather than the previous addition of 100 ml of
soil extract with 900 ml of seawater, 50 ml (1:20), 100 ml (1:10) and 150ml (1:7)
additions of the three extracts were added to 900 ml of seawater. To ensure consistent
salinity, 100 ml and 50ml of rainwater were added to the 50 ml and 100 ml dilutions
respectively. The control consisted of 150 ml of rainwater and 900 ml of seawater.

Bioassay using Lyngbya majuscula from different sites

Again the basic protocol from experiment 1 was replicated. 100 ml additions of cleared
pine and coffee rock extracts were added with either seawater and L. majuscula from the
Deception Bay bloom site or seawater and L. majuscula from the Eastern banks bloom
site.
These three bioassays were conducted for two days each over a fourteen-day period in
January 2001. Prior to addition the extracts were analysed for ammonia, nitrogen oxides,
phosphorus, dissolved and total organic carbon, dissolved and total iron, pH and spectral
properties. L. majuscula from these treatments was analysed for photosynthetic capacity
and photosynthetic pigment (chlorophyll a and phycoerythrin) concentration. During the
incubations the seawater was analysed for ammonia, nitrogen oxides and phosphorus.

Photosynthetic capacity
After 48 hours of incubation a WALZ diving PAM (Pulse Amplitude Modulated)
flourometer was used to measure photosynthetic capacity. Rapid light curves (RLC)
were used to assess the photosynthetic status of the Lyngbya majuscula as described
previously (White & Critchley 1999). Each rapid light curve consists of nine saturating
light pulses (0.8 s) separated by an actinic light. This 10 s actinic light increased stepwise
from 9 to 1667 mol quanta m-2 h-1. At each of these nine steps electron transport rate

- 10 -

(ETR) is calculated and plotted against photosynthetically active radiation (PAR) to

produce the RLC.
ETR = Fv/Fm x PAR x 0.5 x 0.84
Fv = Variable florescence
Fm = Maximum florescence
PAR = Photosynthetically active radiation (umol quanta m-2 s-1)
(White & Chritchley 1999)

Pigment analysis
Both chlorophyll a (chl a) and phycoerythrin (PE) concentrations were assessed after 48
hours of incubation. 0.1g (wet wt.) of L. majuscula tissue was taken and rinsed in filtered
seawater to remove any sediment/particles followed by a rinse in an isotonic solution of
6% ammonium formate to remove excess salts. L. majuscula tissue from each incubation
chamber was sampled for both PE and chl a and frozen immediately.
Phycoerythrin
For PE analysis the L. majuscula tissue samples were ground in a pestle and mortar using
a phosphate buffer solution (pH 6.5, Buffer value 0.024 - 50% 0.01M KH2PO4 13.9%
0.01M NaOH, 36.1% distilled water). This extract was then transferred to a 15ml
centrifuge tube and made up to 10ml with the buffer solution and extracted overnight.
Following centrifugation at 2500 rpm for 20mins, the supernatant was transferred to a
quartz curvette. Pigment determination was then conducted spectrophotometrically using
absorbance at 565 and 750 as indicators of PE concentration and turbidity respectively.
Using specific phycoerythrin formulas (Rowan 1989) phycoerythrin concentration as a
function of dry tissue weight was then determined using a previously calculated Wet
weight : Dry weight ratio.
Chlorophyll a
For Chl a analysis the L. majuscula tissue samples were ground in a pestle and mortar
using 90% Acetone. This extract was then transferred to a 15ml centrifuge tube and
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made up to 10ml with the buffer solution and extracted overnight. Following
centrifugation at 2500 rpm for 20mins then supernatant was transferred to a quartz
curvette. Pigment determination was then conducted spectrophotometrically using
absorbance at 664 and 750 as indicators of chlorophyll a concentration and turbidity
respectively. Using formulas derived from Parsons et al. (1984) concentration as a
function of dry tissue weight was then determined.

Extract Analysis
Soluble and total iron were analysed using inductively couple plasma atomic emission
spectrometry (ICP-AES). Soluble iron is that which passed through a 0.45M GF/F
filter. Extract pH was assessed using a pH probe attached to a 90-FL Field Lab (TPS).
Dissolved and total organic carbon were assessed using by wet oxidation with sodium
persulphate on a total carbon analyser. Seawater samples for nutrient analysis were
filtered through a 0.45 um filter prior to NOx, NH4 and PO4 (FRP) analysis using a
automated LACHAT 8000QC flow injection analyser (FIA).

Spectral Analysis
Water samples were collected from Mellum Creek and an earthern drain in a pine
plantation (sites 9 and 10) both before and after a heavy rainfall event. Waters were
collected in black plastic 20 L drums and kept chilled prior to analysis (within 12 hours).
50 mls of solution was passed through a sartorius 0.45 M syringe mounted filter unit to
remove particulates. Absorbance spectra were then assessed over 300-800nm (1nm
intervals) through a 5cm path length using a Beckman DU500 scanning
spectrophotometer. To remain consistent with previous studies (Kirk 1976, Longstaff et
al. 2001) gilvin 440 readings were converted to a 1m path length.

Statistical analysis
Cochrans test was used to check that variances were homoscedasic. One-way analysis
of variance (ANOVA) was used to test difference in means between treatments. A post
hoc Tukeys test was used to assess which treatments were significantly different.
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Results
Chemical parameters of soil extracts
Extracts derived from the eight various soil types are diverse in both spectral absorbance
and chemical composition (Table 2). This diversity is a function of both soil type and
land use at the sites. The three podosol soil extracts (cleared pine, intact pine and
Melaleuca) all contained significantly more dissolved organic carbon than the other sites.
This was also reflected by high gilvin (absorbance at 440nm (1m path length)) values.
Different land uses within these podosols also showed variation, with the cleared pine
yielding higher dissolved organic carbon than the native Melaleuca forest.
The soil extracts can be categorised into two distinct groups based on pH (Table 2). The
three forested sites (cleared pine, intact pine, Melaleuca) and the canal development all
yielded acidic extracts ranging from pH 3.3 - 4.3. The remainder of the sites yielded
neutral extracts of pH 5.9 - 6.8.
Large variations in free reactive phosphorus (FRP) concentration occurred between the
extracts, cleared pine and intact pine extracts contained 6.7 and 9.5 M P respectively.
The remaining extracts contained negligible FRP concentrations of 0.05 - 0.3 M P
(Table 2).
Extract

pH

Cleared Pine

Mangrove

3.9
3.3
4.3
6.1

ShirleyCreek

5.9

6.2

6.8

Intact Pine
Melaleuca

Sandstone
Point
Canal
Development
Coffee Rock

Gilvin
(g440)
59.5
36.9
36.9
1.9

[PO4]
(M)
6.7
9.5
0.3
<0.01

DOC
mg/L
62.9
35.8
59.1
4.1

TOC
mg/L
1270
1490
255
20

0.2

5.3

470

0.05

43.0

<0.01

0.8

0.1

2.1

20

<0.01

1.0

<0.01

3.7

3.1

0.1

4.7

15

0.15

3.3

0.05

6.6

38.2

<0.01

28.6

240

0.2

Table 2: Chemical parameters of soil extracts.

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Soluble Total Fe Fe(sol):Fe(tot)

Fe mg/L
mg/L
0.57
1.1
0.50
0.12
1.0
0.12
0.43
2.8
0.15
0.16
3.5
0.05

6.1

0.03

Large variation in total iron occurred between the extracts, with Shirley Creek yielding
43 mg/L Fe and others between 0.99 mg/L (intact pine) and 6.1mg/L (coffee rock) (Table
2). The concentration of soluble iron however, did not follow these same trends. Shirley
Creek yielded only 0.05 mg/L of soluble iron, whilst cleared pine extract contained 0.57
mg/L soluble. Fe. When the proportion of soluble:total iron is considered, the three
forested sites (cleared pine, intact pine and Melaleuca) had high proportions of dissolved,
0.50, 0.12 and 0.12 respectively. Other extracts had low ratios of between 0 and 0.05
Soluble:Total Fe. The high ratios of soluble iron in the forested sites generally correlated
with the dissolved organic carbon content of the extracts as well (Figure 4, R2 = 0.78).
0.6

Soluble iron (mg/L)

0.5
0.4

R2 = 0.7834

0.3
0.2
0.1
0
0

10

20

30

40

50

60

70

Disolved organic carbon (mg/L)

Figure 4: The correlation between dissolved organic carbon (DOC) and soluble iron within the eight soil
extracts.

Bioassay results from eight primary land uses

Nutrient Uptake
Uptake rates of phosphorus by Lyngbya majuscula where generally low, with rates of
between 0.01 and 0.06 uM P h-1 in most of the incubation chambers (Table 3). Chambers
with coffee rock, cleared pine and canal development extracts had higher uptake rates of
0.90, 0.17 and 0.13 P uM h-1 respectively (Table 3). Similarly, nitrogen oxides were
assimilated at rates of between 0.05 and 0.17 uM NOx h-1 (Table 3) in most of the
incubation chambers. Chambers with coffee rock, canal development and cleared pine
extracts had elevated uptake rates of 0.51, 0.30 and 0.25 uM NOx h-1 respectively (Table
3). Changes in ammonia concentrations within the incubation chambers were more
- 14 -

variable between treatments. Uptake rates in the seawater control, intact pine, Melaleuca
and mangrove were between 0.12 and 0.21 uM NH4 h-1 (Table 3). Higher rates of 0.39
and 0.66 uM NH4 h-1 occurred in the Sandstone Point and Sandstone+ Pine incubations
respectively. Negligible rates of 0.02 and 0.05 NH4 uM h-1 occurred in response to the
Shirley Creek. and canal development extracts, and release of ammonia occurred in the
cleared pine and coffee rock chambers at rates of 0.04 and 0.47 NH4 uM h-1 respectively
(Table 3).
Extract

Phosphorus
-1
(M h )

Nitrogen Oxides
-1
(M h )

Ammonia
-1
(M h )

Seawater Control

0.05

0.11

0.19

Cleared Pine

0.17

0.25

-0.04

Intact Pine

0.01

0.13

0.12

Melaleuca

0.01

0.05

0.21

Mangrove

0.02

0.11

0.15

Shirley Crk.

0.05

0.17

0.02

Sandstone Pt.

0.06

0.10

0.39

0.13

0.30

0.05

0.90

0.51

-0.47

0.01

0.10

0.66

Canal Dev.
Coffee Rock
Sandstone Pt. +
Pine

Table 3: Phosphorus, nitrogen oxides and ammonia uptake rates from the various incubations over the first
3 hours after Lyngbya majuscula addition.

Photosynthetic capacity
The electron transport rate of Lyngbya majuscula using PAM florescence has been used
to indicate photosynthetic capacity. The L. majuscula in the seawater control had an
electron transport rate of 189 mol m-2 s-1 (Figure 5). The electron transport rate in
response to cleared pine extract was more than doubled (381 mol m-2 s-1), intact pine
showed a 33% increae (252 mol m-2 s-1) and the combination of sandstone point and
pine extracts caused a 74% (329 mol m-2 s-1) elevation (p<0.05) (Figure 5). Depressed
electron transport rates occurred in L. majuscula in response to mangrove, Shirley Creek.,
canal development and coffee rock extracts, although not significant (p>0.05).

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Se
aw
at
er
C
on
tro
C
l
le
ar
ed
Pi
ne
In
ta
ct
Pi
ne
M
el
al
eu
ca
M
an
gr
ov
Sh
e
irl
ey
C
Sa
re
ek
nd
st
o
ne
C
an
Po
al
in
D
t
ev
el
op
m
en
C
t
of
fe
e
Sa
R
oc
nd
k
st
on
e
+
Pi
ne

Photosynthetic capacity
ETR max. (umol m-2 s-1)

450
400
350
300
250
200
150
100
50
0

p<0.05

Soil Extract

Figure 5: Photosynthetic capacity of Lyngbya majuscula incubated in 1:9 dilutions of various soil extracts.

Pigment concentration
The Lyngbya majuscula in the seawater control had 2.7 mg/g DW of phycoerythrin and
2.0 mg/g DW of chlorophyll (Figure 6). All other treatments had trends toward a
reduction in chlorophyll a concentrations, although differences were not statistically
significant (p>0.05). Large variability in phycoerythrin concentrations in L. majuscula
occurred in response to the extracts. All treatments caused an increase in phycoerythrin
concentration, however the only statistically significant (p<0.05) increases, were in
response to cleared pine extract (8.3 mg/g DW), intact pine extract (7.3 mg/g DW) and

12
10
8
6
4
2

ne

Pi

oc

R
e
on

fe
of

st
nd
Sa

D
al
C

an

el
ev

st
nd
Sa

t
m
op

e
on

y
rle
Sh
i

Chlorophyll

en

t
Po

re
C

gr
an
M

Soil Extract

p<0.05

in

ek

e
ov

ca
eu
M

el

al

Pi
ct
ta
In

le

ar

ed

on

Pi

tro

ne

ne

Pigment concentration
(mg/g DW)

coffee rock extract (6.3 mg/g DW) (Figure 6).

Phycoerythrin

Figure 6: Pigment concentrations (chlorophyll a and phycoerythrin) of Lyngbya majuscula incubated in

1:9 dilutions of various soil extracts.

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Bioassay results from various extract dilutions

Nutrient uptake
Uptake rates of phosphorus by L. majuscula where generally low, with rates of between
0.01 and 0.03 uM P h-1 in most of the incubation chambers (Table 4). Chambers with the
1:20, 1:10, 1:7 dilutions of cleared pine extract had higher uptake rates of 0.07, 0.06 and
0.07 P uM h-1 respectively (table 4). Nitrogen oxides in the seawater control were
assimilated at 0.44 uM NOx h-1 (Table 4). The 1:20 dilutions of coffee rock and
sandstone point extract had elevated uptake rates of 0.74 and 0.68 uM NOx h-1
respectively (Table 4). All extracts showed a stepwise decrease in nitrogen oxide uptake
rates with increasing extract concentration. Uptake of ammonia within the incubation
chambers was variable between treatments. A net increase in ammonia concentration
occurred in the seawater control of 0.03 uM NH4 h-1 (Table 4). A stepwise increase in
ammonia uptake rate occurred with increasing concentration of coffee rock and sandstone
point extract, to maximal rates of 4.98 and 2.41 uM NH4 h-1 respectively. Net release of
ammonia occurred in response to 1:10 and 1:7 dilutions of cleared pine extract (Table 4).

Extract

Phosphorus
-1
(M h )

Nitrogen Oxides
-1
(M h )

Ammonia
-1
(M h )

0.01

0.44

-0.03

Cleared Pine 1:20

0.07

0.42

0.25

Cleared Pine 1:10

0.06

0.28

-0.13

Cleared Pine 1:7

0.07

0.25

-0.04

Sandstone Pt. 1:20

0.01

0.74

1.90

Sandstone Pt. 1:10

0.01

0.61

3.03

Sandstone Pt. 1:7

0.03

0.53

4.98

Coffee Rock 1:20

0.01

0.68

1.01

Coffee Rock 1:10

0.01

0.43

1.51

Coffee Rock 1:7

0.03

0.28

2.41

Seawater Control

Table 4: Phosphorus, nitrogen oxides and ammonia uptake rates from the various incubations over the first
3 hours after Lyngbya majuscula addition.

- 17 -

Photosynthetic capacity
The seawater control had an electron transport rate of 197 mol m-2 s-1 (Figure 7), similar
to that of the previous experiment (Figure 7). Electron transport rates were elevated in
the 1:10 and 1:7 dilutions of cleared pine extract (275 and 271 mol m-2 s-1 respectively)
and the combination of sandstone point and pine extracts (380 mol m-2 s-1) (p<0.05)
(Figure 7). Photosynthetic capacity of L. majuscula was not affected by any dilutions of
sandstone point or coffee rock extract.

Photosynthetic capacity
ETR max. (umol m-2 s-1)

450
400
350
300
250
200
150
100
50
0
Seawater
Control

Cleared
Pine 1:20

p<0.05

Cleared
Pine 1:10

Cleared
Pine 1:7

Sandstone Sandstone Sandstone Coffee Rock Coffee Rock Coffee Rock Sandstone
Point 1:20 Point 1:10 Point 1:7
1:20
1:10
1:7
+ Pine
1:1:14

Soil Extract

Figure 7: Photosynthetic capacity of Lyngbya majuscula incubated in 1:20, 1:10 and 1:7 dilutions of
cleared pine, sandstone point and coffee rock soil extracts.

Pigment concentration
Lyngbya majuscula in the seawater control had phycoerythrin concentrations of 3.8mg/g
DW and 1.5 mg/g DW of chlorophyll (Figure 8). Phycoerythrin concentrations were
significantly increased (9-13 mg/g DW) in all dilutions of cleared pine extract (p<0.05).
All dilutions of coffee rock extract also caused a significant increase in phycoerythrin
concentrations, however these were significantly less than that of the cleared pine extract
(p<0.05) (Figure 8). Dilutions of sandstone point had no significant affect on
phycoerythrin concentrations in L. majuscula. The combination of cleared pine and
- 18 -

sandstone point extracts elevated phycoerythrin concentration to 10.6 mg/g DW (Figure

8). Chlorophyll a concentrations were not significantly altered by addition of any of the
various soil extracts (Figure 8).

Pigment Concentration (mg/g DW)

16
14
12
10
8
6
4
2
0
S eaw ater
C ontrol

C leared
P ine 1 :2 0

C leared
P ine 1 :1 0

C leared
P ine 1 :7

S and ston e S and ston e S and ston e

P oint 1:2 0 P oint 1:1 0
P oint :7

C offee
R ock 1:2 0

C offee
R ock 1:1 0

C offee
R ock 1:7

S and ston e
+ P in e
1 :1 :14

S oil Extract/D ilution

p<0.05

C hlo rop hyll

P hycoe rythrin

Figure 8: Pigment concentration (chlorophyll a and phycoerythrin) of Lyngbya majuscula incubated in

1:20, 1:10 and 1:7 dilutions of cleared pine, sandstone point and coffee rock soil extracts.

Bioassay results using Lyngbya majuscula from different sites

Photosynthetic capacity
The photosynthetic capacity of Lyngbya majuscula sampled from Deception Bay and
Eastern Banks was increased by addition of cleared pine extract but not affected by
coffee rock extract (p<0.05). These results were consistent with previous experiments
(Figure 9).

- 19 -

(ETR max umol m-2 s-1)

Photosynthetic capacity

Deception
Bay

Eastern
Banks

350
300
250
200
150
100
50
0
Seawater
Control

Cleared
Pine

Coffee
Rock

Seawater
Control

Cleared
Pine

Coffee
Rock

Soil Extract

p<0.05

Figure 9: Photosynthetic capacity of Lyngbya majuscula from the Eastern Banks and Deception Bay
bloom sites incubated in 1:9 dilutions of cleared pine and coffee rock soil extracts.

Pigment concentration
Phycoerythrin concentration of Lyngbya majuscla from Deception Bay and Eastern
Banks was elevated by additions of coffee rock and cleared pine extracts (Figure 10).
The treatments and controls from Deception Bay had significantly (p<0.05) higher
concentrations of phycoerythrin than that of the Eastern Banks. No change in chlorophyll
a concentrations were observed between treatments or sites (Figure 10). Therefore the

Pigmnet concentration (mg/g

DW)

two L. majuscula populations appear to be physiologically similar.

Eastern
Banks

12.0
10.0

Deception
Bay

8.0
6.0
4.0
2.0
0.0
Seawater
Control

Cleared
Pine

Coffee
Rock

Seawater
Control

Cleared
Pine

Coffee
Rock

Soil Extract

p<0.05

Chlorophyll

Phycoerythrin

Figure 10: Pigment concentration (chlorophyll a and phycoerythrin) of Lyngbya majuscula from the
Eastern Banks and Deception Bay bloom sites incubated in 1:9 dilutions of cleared pine and coffee rock
soil extracts.

- 20 -

Spectral properties of waters

The absorbance of creek waters sampled from the Pumicestone region followed distinct
patterns across the UV-VIS spectrum. The UV(B) region (280-315 nm) shows variable
absorbencies between 0.02 and 0.8. The UV(A) region (315-400 nm) was dominated by
a large peak at 295 nm (Figure 11). From 330-750 the absorbance gradually declined. It
is likely that the peak at 390 nm is an aberration of the transition from VIS-UV light
sources within the spectrophotometer. The pine drain had absorbencies twice that of
Mellum Creek across the 400-600 nm range in both dry and wet periods. In both creeks,
there was also a two-fold increase in absorbencies during the high rainfall (wet period)
compared to the low rainfall (dry) period (Figure 11).

Absorbance (5cm path length)

3.5
3
2.5
Mellum Crk. (Dry)

Pine Crk. (Wet)

Pine Crk. (Dry)

1.5

Mellum Crk. (Wet)

1
0.5
0
200

300

400

500

600

700

800

Wavelength (nm)

Figure 11: Spectral absorbance (5cm path length) of Mellum Creek and a pine drain before and after a
heavy rainfall event.

- 21 -

Discussion
Blooms of Lyngbya majuscula have been increasing in the Deception Bay region of
Moreton Bay region over recent years. It is hypothesised that altered runoff dynamics
due to land use change within the catchment may have contributed to these increases.
Runoff during rain events may be releasing iron, phosphorus and dissolved organics,
which have been shown in other regions to stimulate algal blooms (Bennet et al. 1986,
Mallin et al. 1991, 1993). The Pumicestone catchment, which leads into Deception Bay,
contains a diversity of horticultural, residential and natural land uses. The dominant land
use within the catchment is exotic pine plantations (Pinus elliottii) (39%), a third of
which have been clear-felled in the last decade (Figure 12). This current study has
assessed the potential for these various land uses within the Pumicestone catchment to be
the source of substances stimulating blooms of L. majuscula in Deception Bay.
Large variability occurred in the chemical composition of soil extracts derived from the
different land uses. Due to the high capacity for forests to fix atmospheric carbon the
cleared pine, intact pine and Melaleuca forest soils yielded higher organic carbon than
un-forested sites. Differences between forests however are more compelling. Total
organic carbon was higher in the pine forests compared with that of the Melaleuca
forests. These differences have been observed previously over a twelve-week leaching
study, pine plantations yielded twice the organic carbon as that of a natural oak forest,
and four times that of grasslands (Khomutova et al. 2000). These differences are likely a
product of the lower C: N ratio of pine litter and higher surface area: volume ratio
enabling rapid microbial incorporation into the soil.
Phosphorus generally has a strong affinity for binding to particles, it is generally
considered immobile within soils, and attached to suspended sediment particles in runoff
(Johnson et al. 1976, Duffy et al. 1978). Therefore, the high dissolved phosphorus
concentrations within the cleared and intact pine forest extracts would not generally be
anticipated. However, recent research has shown that in forested catchments phosphorus
is predominately attached to materials <0.45 M (solubilised) (Sharpley et al. 1994, Nash
& Murdoch 1997). This may be a result of phosphorus being able to bind to Fe-organic
complexes making it transportable (Francko & Heath 1983). Similar to the
- 22 -

photoreduction of Fe(II) from these complexes, UV light is also able to drive the release
of orthophosphate from the P-Fe-organic complex (Francko & Heath 1983, Cotner &
Heath 1990). However in the absence of Fe(III), phosphorus shows little affinity for
forming a soluble P-organic complex (Koenings and Hooper 1976).
The forested soils (pine and Melaleuca) yielded acidic extracts between pH 3.5-4.3. This
acidification of forested extracts has been noted previously in leaching trials using intact
soil columns of pine and grassland sites (pH 3.6 and 7.1 respectively) (Khomutova et al.
2000). This acidification allows iron oxides and phosphates present within the soil to
become mobilised. This mobilised iron, in combination with the high organic matter of
these forested soils, would likely yield an iron-organic complex.

Relationship between soluble iron and dissolved organic carbon

In the current study, there was a positive correlation between the soluble iron
concentrations and the organic matter contained in the different soil extracts. Previous
studies have shown that organic carbon from both terrestrial and marine sources is able to
complex with Fe(II) or Fe(III) (Theis and Singer 1974, Koenings & Hooper 1976, Morel
1983). Extracts without high concentrations of organic material, such as Shirley Creek,
had very high total iron but none was present in the soluble phase. Whereas, organic rich
extracts did not necessarily have high total iron content, but a high proportion of what
was there was in the soluble phase. The coffee rock extract was the only exception to this
trend, having high dissolved organic carbon with negligible iron present in the soluble
phase. This is likely due to the difference in the specific organic compounds between the
forested soils and coffee rock. Specific organics types have differing abilities to complex
iron (Hutchins et al. 1999). As only DOC and TOC assessments were made of these
extracts, it is beyond the scope of this study to draw any links between soluble iron and
specific organic types (e.g. fulvic acid etc.). Parallel studies, have indicated that the
organics within the pine extracts are able to complex the iron 240 times the rate of the
organics in the coffee rock extract (Rose unpub. data). Thus, not only are pine forests
yielding more organic rich material into Deception Bay, that organic material is able to
complex iron far more effectively than organics from the native vegetation.

- 23 -

The acidity of the forest soils shifts the thermodynamic preference towards soluble ferric
iron. Coupled with this, acidification of soils reduces their capacity to absorb organic
matter and hence yields a greater dissolved organic carbon component (Forsberg 1992).
These factors would contribute to the elevated soluble iron in the acidic forest soil
extracts. However, there was no direct correlation (R2 = 0.0816) between soluble iron
and pH. Cultivation of pine forest soils also increases organically bound iron and
aluminium (Dormaar 1979, Zhang 1988). It has been previously noted that soils under
coniferous species such as Pinus elliottii generally have high concentrations of
organically bound iron and aluminium (Quideau & Bockheim 1996, Khomutova 2000).
This may be a function of the three above mentioned factors (DOC, pH and cultivation).
Being soluble, these fine colloidal organic-iron complexes may be transported in the
water column. Prior to release of free Fe(II)/Fe(III) a process such as photo-reduction
must occur (Voelker et al. 1997, Waite & Morel 1984, Wells & Mayer 1991). This is
somewhat similar to what has been hypothesised to occur in Deception Bay. The
organics from the pine provide a transport mechanism for the iron to reach the bloom site,
upon reaching the shallow high light environment photo-reduction occurs, releasing bioavailable iron to the L. majuscula. Similar patterns of iron/organic dynamics have been
noted in Swiss freshwater lakes (Emmenegger et al. 1998), where cyanobacterial blooms
have been linked with terrestrial organics providing a source of bioavailable iron.

Soil extract effects on photosynthetic capacity of Lyngbya majuscla

The main focus of this study was to investigate how the soil extracts (from different land
uses) affected physiological parameters of Lyngbya majuscula. A significant (p<0.05)
increase in photosynthetic capacity of L. majuscula occurred in response to extracts from
cleared and intact pine forests. Both the cleared and intact pine extracts can be distinctly
separated from the others by their high phosphorus concentrations (6.7 & 9.5 M
respectively). It is unlikely that the elevated photosynthetic rates observed in response to
these extracts are a result of phosphorus alone, as the cleared pine extract caused a
significantly higher (p<0.05) photosynthetic response compared with that of the intact
pine extract, yet had a significantly lower phosphorus concentration. The cleared pine,
however, did contain soluble iron concentrations four-fold those of the intact pine
- 24 -

extracts. It appears, based on these results, that a threshold concentration of phosphorus

may be required for elevation of the photosynthetic capacity of L. majuscula. The
magnitude of this elevation however, may be related to soluble iron concentration not
phosphorus. This pattern is re-enforced by the results obtained from L. majuscula treated
with the other extracts. The Melaleuca, mangrove, canal development and coffee rock
extracts all contained significantly higher soluble iron than the intact pine extract, yet,
unlike the intact pine, the extracts from these soils contained low phosphorus
concentrations, and hence lacked the photosynthetic response.
These incubations indicate that a combination of iron and phosphorus likely stimulated
the photosynthetic capacity in Lyngbya majuscula. These results correlate with studies of
other cyanobacteria and plankton species in which interactions between iron and
phosphorus were required for elevations in photosynthetic rates (Lovstad & Krogstad
2001, Clasen & Bernhard 1974). However, at the Deception Bay bloom site it is
probable that sufficient iron and phosphorus is present in the sediment to support L.
majuscula growth and terrestrial sources may not be necessarily important. Thus, it is the
dissolved organic carbon which may become the critical component in complexing the
iron and potentially phosphorus already present on site in the sediment, into the soluble
phase for assimilation by L. majuscula.

Nutrient uptake
The Lyngbya majuscula used, was sampled from waters with very low dissolved [PO4]
and [NH4] (Watkinson 2000). However, when incubated in elevated concentrations, the
uptake of nitrogen (NH4/NOx) and phosphorous was extremely rapid, with the bulk
absorbed within the first hour. It is interesting that L. majuscula appears to be
assimilating ammonia and nitrogen oxides despite being capable of nitrogen fixation.
During late stages of Lyngbya majuscula blooms it has been previously observed that
nitrogen fixation rates decrease compared to earlier stages of the bloom (Duffy & ONeil
submitted). Thus, it would be expected alternate nitrogen sources such as ammonia
would be sought. Indeed, nitrogen fixing cyanobacteria can assimilate ammonia directly
from the water column, particularly during late stages of the growth cycle (Mulholland &
Capone 2000), potentially due to the metabolic costs of nitrogen fixation.
- 25 -

With increasing concentrations of extract added, the uptake of ammonia increased

concomitantly. This uptake of excess nutrients has been observed previously in
phytoplankton (McCarthy & Goldman 1979). Hypothetically, if nutrient runoff into
Deception Bay were to increase, Lyngbya majuscula would have the capacity to rapidly
assimilate this and potentially increase growth rates.
Land use affects catchment hydrology
The potential for elevated terrestrial nutrient run-off into Deception Bay waters is high,
with large changes in catchment land use in recent years. The Pumicestone catchment
contains a diversity of horticultural, residential and natural land uses. The dominant land
use within the Pumicestone catchment is exotic pine plantations (Pinus elliottii) (39%).
Plantations of exotic P. elliottii within SE Queensland are grown as a monoculture,
predominantly in coastal lowland regions dominated by low nutrient sandy podosol soil.
Within the last 5 years, a third of the total plantation area has been clear-felled, partly as a
result of salvage logging after bushfires in 1995-96 (Figure 12).

Figure 12: Areas of pine plantation clear-felled within the Pumicestone catchment over the last 12 years.
Source: Christian Witte Department of Natural Resources and Mines, Queensland Government.

- 26 -

The conversion of conventional crops (e.g. sugar cane, orchards) to short rotation woody
plantations (e.g. Pinus sp.) reduces eutrophication of waters and reduces the quantity of
runoff, due to the reduced fertilizer requirements and deeper root penetration of woody
plantations (Joslin et al. 1997). However, the Joslin (1997) study was based during the
growth phase of the plantations and did not consider the impacts of harvesting on water
quality and quantity. Large-scale clear-felling of forests has been shown to elevate
dissolved nutrient concentrations, suspended sediment loadings and net run-off volume
into surrounding waters (Cambell & Doeg 1989).
Mobilisation of organics from forested areas is enhanced following sufficient rainfall to
saturate the soil column. The high evapotranspiration rate within forests is generally
sufficient to maintain the water table below the surface, and hence prevent excess run-off.
However, due to the clear-felling of pine plantations this hydrological balance is upset
such that elevated run-off of organics occurs. Following clear-felling in the Pumicestone
catchment, darkly stained, organic rich waters have been observed entering Pumicestone
passage (Figure 13). It can take 4-8 years after clearing for evapotranspiration to become
sufficient enough to lower the water table (Bubb pers comm.). It is likely that large scale
clearing of any vegetation type within the region will cause similar hydrological changes.

Figure 13: Organic rich waters entering Pumicestone Passage through

Westaways Creek (Bribie Is.), following pine plantation clear-felling.

- 27 -

Organic rich run-off into coastal waters

Algal blooms in estuaries worldwide have in many cases been linked to terrestrial runoff, following periods of heavy rainfall (Bennet et al. 1986, Mallin et al. 1991, 1993).
More specifically, the introduction of dissolved organic matter as a result of this rainfall
has been attributed to phytoplankton blooms (Heil 1996). High levels of dissolved
organic matter are common in the coastal waters of Australia, resulting in stained water
colour in some regions (Kirk 1994, Figure 13). Evidence exists, that the input of organic
material into natural waters has been increasing in some regions across Europe over the
last 15 years (Andersson et al. 1989, Forsberg and Peterson 1990).

Spectral data on creeks entering Pumicestone Passage indicate that high organic loadings
enter the passage from both mainland and Bribie Is. areas. Total organic carbon within
creeks adjacent to pine forests can reach levels up to 28.5 mg/L (Watkinson 2000), far in
excess of background levels. Distinct increases in organic absorbances occurred in
Mellum Creek and a pine drain following a rainfall event in early February. During the
high rainfall period both creeks had gilvin (absorbance at 440nm) readings higher than
that of previous studies of Australian water bodies (Kirk 1976). As explained previously,
heavy rainfall sufficient to saturate the soil column yields high concentration of organics
in the soluble phase.

Long term leaching studies of soil cores from pine forests have shown that after 20
weeks, 50% of the organic carbon is removed through rainfall leaching (Khomutova et al.
2000). After that period leaching slowed considerably. Although conditions may vary in
actual forests, it can be expected that following clear-felling, the majority of organic
carbon will be leached within the first 20-30 weeks. Based on this evidence, potential
management strategies are able to be recommended. One possibility for instance, is that
clear-felling of large areas of pine plantation be conducted in late summer after the
Lyngbya majuscula blooms and heavy rainfall periods have subsided. This would allow
the following 20-30 weeks for much of the organic carbon to leach out during the winter
months, when waters are colder, without the threat of promoting L. majuscula blooms.
By the following summer, when the waters are warm enough for L. majuscula to bloom,
- 28 -

a cover crop with high transpiration capability could have been planted on the clear-felled
areas, increasing evapotranspiration and further reducing the leaching of remaining
organic matter. Further lab based leaching studies have yielded higher organic carbon
leaching with increasing air temperature (Duffy et al. 1989). This may further compound
the elevated organic carbon leaching into Pumicestone Passage during the warm summer
months.

Dissolved organic carbon effects light quality

Coupled with the iron complexation dynamics, dissolved organic carbon has also been
attributed to a reduction in light quality within the water column. The spectra of sunlight
can be generically divided into ultraviolet (UV-200-400nm), blue (400-500 nm), green
(500-600 nm) and red (600-750 nm) bands. Absorbance spectra of extracts from this
study show high absorbance in blue region of the spectrum. Organics have been
previously shown to absorb primarily in the UV and blue (400-500 nm) regions of the
spectra (Kirk 1976). The parameter gilvin (absorbance at 440nm) has been coined to
characterize the absorbance of these organics. Seawater has a background absorbance of
red light. Thus in seawater containing organic material the majority of light available to
organism is green (500-600 nm). Chlorophyll a has a primary absorbance peak in the
blue band (Soret band) and a secondary peak in the red band, hence, it is unable to absorb
sufficient light in organic rich waters. L. majuscula, like many cyanobacteria, has a
specialized pigment (phycoerythrin), which has an absorption maxim at 565 nm (green).
This may be an evolutionary adaptation to having to inhabit organic rich waters in order
to receive sufficient iron, giving it a preferential advantage over flora reliant on
chlorophyll a. The impacts of dissolved organics on both chromatic quantity and quality
has been observed previously (Jerlov 1955, Kirk 1976, Grantham 1981, Doering et al.
1994, Kirk 1994, Davies-Colley et al. 1993, Schwarz & Markager 1999)

- 29 -

Changes in light quality effect cyanobacterial pigments

Red algae (Rhodophyta) are able to increase either chlorophyll or phycoerythrin
production based on the light environment exposed to (Aguirre-von-Wobeser et al. 2001).
Rapid changes in pigment ratios over diurnal cycles in response to changes in light
quality have also been observed (Lopez 1992). More specifically, light quality can
influence photosynthetic pigments and productivity in Lyngbya majuscula (Longstaff et
al. 2001). Phycoerythrin concentrations in L. majuscula increased following alteration
of ambient light quality similar to that experienced in Deception Bay as a result of
dissolved organics in the water column. In the current study, a strong correlation was
found between gilvin readings in the extracts and the phycoerythrin concentration of the
L. majuscula. The only exception to this correlation was the lowest addition of cleared
pine extract, which caused high phycoerythrin concentrations despite relatively low
gilvin levels. This anomaly may be due to the fact that the pine extract had high amounts
of soluble iron for phycoerythrin production and was, hence, less controlled by light
quality. This general trend of high phycoerythrin concentrations in water with high gilvin
is supported by comparisons of the Deception Bay and Eastern Banks L. majuscula
controls. Without addition of extracts, the L. majuscula from Deception Bay had
significantly higher phycoerythrin concentrations than that of L. majuscula from the
Eastern Banks, most probably as a result of the spectral differences in the environment
where each had been acclimatised.
Elevated phycoerythrin concentrations of L. majuscula in response to cleared pine
extracts may be enabling the increase in photosynthetic capacity observed due to greater
photon capturing ability. The coffee rock treatment however, had elevated phycoerythrin
concentrations without any apparent effect on photosynthetic capacity. Similar increases
of phycobillins in the pelagic cyanobacterium Trichodesmium have been observed
without subsequent photosynthetic increases (Trick et al. 1995). It was hypothesised that
the phycobillins in this instance, were in an inactive state and used only for nitrogen
storage. Previous studies indicate that iron availability can also influence phycoerythrin
concentrations (Sandmann 1985). However, in the current study, the correlation between
soluble iron and phycoerythrin was not evident (R2=0.169).

- 30 -

Conceptual summary of four extract groupings

(a)

(b)

(c)

(d)

Figure 14(a,b,c,d): Conceptualisation of interactions between soil extracts, light and Lyngbya majuscula
physiology (relative symbol sizes reflect concentrations of various parameters).

- 31 -

The data derived from this study can be summarized into four distinct groupings based on
land use.
Lyngbya majuscula treated with extracts derived from Sandstone Point and Shirley Creek
sediments received the full sunlight spectrum and thus had equal ratios of phycoerythrin
and chlorophyll a. Negligible soluble iron was present in the extracts due to the lack of
organics. This lack of available iron combined with low phosphorus concentrations
limited photosynthetic activity (Figure 14a).
Lyngbya majuscula treated with extracts derived from mangrove and canal development
soils received the full sunlight spectrum and thus had equal ratios of phycoerythrin and
chlorophyll a. The majority of iron was not soluble due to the low organics present. This
low available iron combined with low phosphorus concentrations resulted in limited
photosynthetic activity (Figure 14b).
Lyngbya majuscula treated with coffee rock and Melaleuca extracts received a reduced
spectrum of light due to the dissolved organics filtering out the blue region. This caused
an elevation of the phycoerythrin concentrations relative to chlorophyll. The dissolved
organics complexed some iron into the soluble phase however phosphorus was not
present thus, photosynthetic activity was limited (Figure 14c).
Lyngbya majuscula treated with cleared and intact pine extracts received a reduced
spectrum of light due to the dissolved organics filtering out the blue region. This caused
an elevation of the phycoerythrin concentrations relative to chlorophyll. Dissolved
organics complexed large amounts of iron into the soluble phase. High concentrations of
phosphorus coupled with this available iron caused an elevation in photosynthetic
capacity (Figure 14d). Coupled with the potential for pine forest soils to stimulate L.
majuscula growth, are the increased loadings of run-off from pine forests into Deception
Bay, as a result of hydrological impacts associated with clear-felling.

- 32 -

Global Implications
The combination of factors required for blooms of Lyngbya majuscula to occur is
specific: high concentrations of iron and phosphorus in either the sediment or water
column, dissolved organics in water column, warm waters and shallow seagrass beds. It
is hypothesized that increases in dissolved organics in the water column has provided the
missing link for enabling the bio-availability of the iron and phosphorus present.
Similarly, other blooms of cyanobacteria have been definitively linked with terrestrial
organics providing a source of bioavailable iron (Emmenegger et al. 1998). Other L.
majuscula bloom areas in the world may potentially have similar organic sources.
Bamboo and sword-grass trash is seasonally flushed into coastal waters of Guam during
the summer months of the L. majuscula blooms (Matson 1991) although the link has not
been specifically drawn. The recent seasonal blooms of L. majuscula in Florida (USA)
are in the proximity of large freshwater Melaleuca swamps which yield similar dark
stained waters to those observed in this study (Burns pers. comm.). Red tides
(dinoflagellate blooms) in Florida have also been linked with dissolved organics in the
water column (Ingle & Martin 1971).
It has been hypothesised that the greenhouse effect may be causing elevated primary
productivity in forests, which in turn is driving long-term trends of increasing dissolved
organic carbon and subsequent increases of water colour in natural water bodies
(Forsberg 1992, Doering et al. 1994). Based on this, in the coming decades coastal
waters may become warmer, more coloured and high in soluble complexed iron,
providing a hospitable environment for cyanobacteria. However, in some cases longterm trends indicate dissolved organic carbon inputs into the coastal marine system are
decreasing. Urbanisation of catchments and associated deforestation has reduced forest
derived organic matter loadings (Kawaguchi et al. 1997, Matsunaga et al. 1999). The
resulting reduction in iron availability caused significant changes in the marine floral
ecosystem in both studies. These studies were conducted in catchments that had been
urbanized for some time. In contrast, the present study has focused on the short-term
elevation of organic matter loadings and elevated iron availability following
deforestation. Eventually the Pumicestone catchment may be more urbanized, which will
potentially result in a reduction in the bio-available iron, with further, yet unknown
- 33 -

impacts on the marine system. Irrespective of the long-term trends in organic carbon
loadings, it is apparent that forests provide a critical function in regulating iron
availability in coastal waters. Anthropogenic manipulation of coastal forests can upset
the iron balance and, in turn, impact on the ecological integrity of the marine system.
The Lyngbya majuscula blooms in this study provide an example of this. Based on this
relationship between catchments and iron dynamics within the associated estuary,
Kawaguchi et al. (1994) have proposed bio-available iron as an indicator of upstream
catchment health.

Conclusions
It is concluded by this study that the changes in catchment land use can increase the
terrestrial nutrient input into coastal marine systems, altering water chemistry and in turn
leading to subsequent impacts on ecological integrity (e.g. formation of nuisance
cyanobacterial blooms). The results of this study clearly show that pine forest soils are
distinct from the other soils in most of the parameters assessed. The pine soils had higher
organic carbon content, high phosphorus concentration, high soluble iron, low pH, and
caused Lyngbya majuscula to have elevated photosynthetic capacity and high
phycoerythrin concentrations. Specifically, iron and phosphorus concentrations were
correlated with photosynthetic increases. Dissolved organic carbon within the forested
extracts altered light quality that the L. majuscula received. Due to this altered light
spectrum, a shift in the relative proportion of photosynthetic pigments present occurred,
with a shift from chlorophyll a to phycoerythrin. It is likely that this physiological
plasticity in response to environmental parameters, exhibited by L. majuscula, offers a
selective advantage over other marine flora and enables the rapid growth to bloom
proportions seen in the region.
Hydrological changes, associated with pine forest clear-felling, increase the flux of
organics into surrounding waters. Thus, the elevated growth of Lyngbya majuscula in
response to pine forest extracts is potentially exacerbated by more of these compounds
entering the bloom site. The conclusions drawn by this study have direct management
implications for potentially reducing the role terrestrial inputs have in stimulation of L.
majuscula blooms within Deception Bay. Management authorities often consider
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terrestrial and marine systems separately. This study has clearly shown but a few of the
important links, which exist between these systems. These links demonstrate that
terrestrial and marine systems are co-dependant. For the maintenance of ecological
integrity of either system, management must be considered on a holistic basis.

Notes on Methodology
Data from further studies using this methodology has been omitted from this report as the timing
of the bioassays caused discrepancies in the results. The data within this report is based on
bioassays conducted during the peak of the bloom (Jan/Feb) when the L. majuscula is
presumably at a physiological optimum. Bioassays conducted in the following months of the
bloom decline showed few significant responses to a large array of treatments. This observation
may lend weight to the argument that bloom decline is virally mediated (Hewson et al 2001),
rather than being a direct physiological response to environmental parameters such as light,
temperature and nutrients. Thus, recommendations for future use of this methodology are that
incubations using a seasonally blooming organism are conducted during the peak bloom period.

Acknowledgments
This research is the result of inputs from a team of people who have taken interest from the first
brainstorming session to the last correction in this thesis. Thanks to Judy ONeil for all her help
during the endless time in the lab and field. Bill Dennison provided much of the guidance and
insight to make this project possible from the outset. Thanks also to Phil Moody and David Waite
who guided me (an ecologist) through difficulties with soil science and iron chemistry respectively.
Although easily convinced, Dan Wruck did the impossible and temporarily relocated his nutrient
analysis lab to our island research station to provide instant analysis. Thanks also to Alan
Goldizen, Andrew Watkinson and all the MarBot crew for lending a hand whenever needed.

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