Aquatic Botany, 24 ( 1 9 8 6 ) 3 5 5 - - 3 7 1 355

Elsevier Science P u b l i s h e r s B.V., A m s t e r d a m - - P r i n t e d in T h e N e t h e r l a n d s

THE LOSS OF SEAGRASS IN COCKBURN SOUND, WESTERN

AUSTRALIA.
III. THE EFFECT OF EPIPHYTES ON PRODUCTIVITY OF
POSIDONIA A USTRALIS HOOK. F.

K. S I L B E R S T E I N ' , A.W. C H I F F I N G S 2 a n d A.J. M c C O M B

Botany Department, University o f Western Australia, Nedlands W.A. 6009 (Australia)
( A c c e p t e d for p u b l i c a t i o n 25 F e b r u a r y 1 9 8 6 )

ABSTRACT

S i l b e r s t e i n , K., Chiffings, A.W. a n d M c C o m b , A.J., 1 9 8 6 . T h e loss o f seagrass in C o c k b u r n

S o u n d , W e s t e r n Australia. III. T h e e f f e c t o f e p i p h y t e s o n p r o d u c t i v i t y o f Posidonia
australis H o o k . f. Aquat. Bot., 24: 3 5 5 - - 3 7 1 .

T h e h y p o t h e s i s was e x a m i n e d t h a t i n c r e a s e d e p i p h y t e g r o w t h was r e s p o n s i b l e for a

r e d u c t i o n in seagrass m e a d o w s in C o c k b u r n S o u n d d u r i n g t h e d i s c h a r g e o f n u t r i e n t -
rich e f f l u e n t . O n e s t u d y site was in a d e t e r i o r a t i n g m e a d o w n e a r a n e f f l u e n t o u t f a l l , t h e
o t h e r at similar d e p t h in a n u n a f f e c t e d m e a d o w in m o r e o c e a n i c w a t e r . Seagrass p r o d u c -
t i o n at t h e first site was less t h a n t h a t a t t h e s e c o n d , w i t h 33% l o w e r g r o w t h per s h o o t
a n d 29% less d e n s e m e a d o w . W a t e r at t h e f o r m e r site h a d h i g h e r m e a n c o n c e n t r a t i o n s
o f c h l o r o p h y l l a n d p h o s p h a t e t h a n t h e l a t t e r , b u t l i g h t r e a c h i n g t h e seagrass m e a d o w s was
n o t s i g n i f i c a n t l y d i f f e r e n t . E p i p h y t e loads (as d r y w e i g h t or c h l o r o p h y l l per u n i t leaf
a r e a ) w e r e 2--8 t i m e s h i g h e r at t h e f o r m e r site. S e a s o n a l c h a n g e s in e p i p h y t e l o a d s w e r e
well c o r r e l a t e d w i t h p e r i p h y t o n b i o m a s s o n glass slides o r plastic seagrass.
P h o t o s y n t h e s i s of leaf s e g m e n t s , w i t h a n d w i t h o u t e p i p h y t e s , was m e a s u r e d u s i n g a n
o x y g e n m e t e r in t h e l a b o r a t o r y ; e p i p h y t e p h o t o s y n t h e t i c r a t e s w e r e similar t o t h o s e o f
p e r i p h y t o n o n plastic, e x p r e s s e d p e r u n i t c h l o r o p h y l l . T h e p e r c e n t a g e r e d u c t i o n in light
b y k n o w n p e r i p h y t o n loads was m e a s u r e d , a n d u s e d t o c a l c u l a t e light r e d u c t i o n b y epi-
p h y t e s in t h e field, w h i c h was e s t i m a t e d t o b e 63% o n average at t h e first site a n d 15%
at t h e s e c o n d . P o o l i n g d a t a for sites a n d seasons, t h e r e w a s a n e g a t i v e log-linear r e l a t i o n -
ship b e t w e e n leaf p r o d u c t i o n a n d e p i p h y t e load. T h e o b s e r v a t i o n s p r o v i d e s u p p o r t for
t h e suggestion t h a t seagrass loss in t h e S o u n d m a y b e a t t r i b u t e d t o e n h a n c e d e p i p h y t e
loads f o l l o w i n g n u t r i e n t e n r i c h m e n t .

INTRODUCTION

Circumstantial evidence has been presented that, of several hypotheses

which may be put forward to explain the loss of seagrass meadows from

1Present address: B o t a n y D e p a r t m e n t , M o n a s h U n i v e r s i t y , V i c t o r i a 3 1 6 8 .
2Also a t D e p a r t m e n t o f C o n s e r v a t i o n a n d E n v i r o n m e n t , 1 M o u n t S t r e e t , P e r t h 6 0 0 0 ,
W e s t e r n Australia.

0304-3770/86/$03.50 © 1 9 8 6 Elsevier S c i e n c e P u b l i s h e r s B.V.

356

Cockburn Sound, Western Australia, the most acceptable one is t h a t en-

hanced growth of epiphytes in nutrient-enriched waters has reduced the
a m o u n t of light available for seagrass photosynthesis (Cambridge et al.,
1984, 1986). The evidence comes from: (1) observations of the levels of
both epiphytes and loosely-attached, blanketing layers of filamentous
algae associated with deteriorating meadows; (2) the coincidence of wide-
spread loss of seagrass with the discharge into the Sound of effluents rich in
plant nutrients; (3) the observed prolific growth of epiphytes 6n leaves of
seagrasses transplanted into the Sound; and (4) the k n o w n importance of
epiphytes in other systems. Other proposed mechanisms could n o t satisfac-
torily account for the general decline in seagrasses, although t h e y m a y be of
supplementary importance in some instances, in that they have brought
a b o u t local decline (as with scouring or oil refinery effluents) or increased
the rate of decline (as with sea urchin grazing and reduction of light trans-
mittance by p h y t o p l a n k t o n blooms).
The work described in this paper was designed to obtain more direct
evidence about the role of epiphytes, and proceeded along several lines. Two
sites were selected in the Sound, one where meadows were declining, the
other where meadows were apparently unaffected. Epiphyte loads and
seagrass p r o d u c t i o n rates were measured for several months, to find o u t if
there was any relationship between the two. Then, as it might be argued that
increased growth of epiphytes could be a consequence rather than a cause
of physiological deterioration in seagrass leaves, measurements were made
of the growth of 'epiphytes' (periphyton) on artificial substrates at the two
localities. Finally, more direct investigations were carried o u t into the
a m o u n t and significance of shading brought a b o u t by epiphytes. The relation
between r a t e of seagrass p h o t o s y n t h e s i s and light intensity was measured
and, through their control over light intensity, the effects of k n o w n loads
of epiphytes on photosynthesis of the seagrass were determined.

MATERIALS AND METHODS

Study sites and measurement o f seagrass growth

Investigations were carried o u t in meadows of Posidonia australis Hook. f.

on the barrier bank at the n o r t h e r n end of Cockburn Sound (Fig. 1). The
first site was towards the eastern end of the bank, close to the o u t l e t of a
sewage treatment plant near W o o d m a n Point. Aerial p h o t o g r a p h y confirmed
that the area and density of seagrass meadows had been reduced between
1977 and 1980. The second site was of the same depth (3 m), but towards
the western end of the bank, near Carnac Island, where water is predom-
inantly oceanic {Chiffings, 1979; Steedman and Craig, 1983).
Productivity was measured essentially by the technique of Zieman (1974).
Holes 1--2 mm in diameter were p u n c h e d through the upper sheath and en-
closed leaf blades with surgical tongue forceps (Kirkman and Reid, 1979),
 357

~ o ~ o...........
0.5 .

.),', .,,,; .... <"....

/7 - I
• ~ FocmIf Seagresa Areas
~//~'/'/////JPcmsenl SiIqirass Areas
We$1em d
Dredged Areas
ALISITIIIa • Sampling Site
. . - - Sewage Ouifall
COCKBURN ~ 5 - - Oeplh Contours In' M@tres
SOUND ~]+ Beacons 10 2 km

Fig. 1. The distribution of seagrass on Parmelia Bank, at the northern end of Cockburn
Sound, showing the distribution of meadows in 1978 and at the time of the present
study, in 1980--81. Study sites 1 (near Woodman Point) and 2 (near Carnac Island) are
shown.

avoiding damage to t he lower sheath. Sho ot s were m arked within seven

plastic-coated steel quadrats, each 0.1 m 2 . T he shoots were harvested after
a p p r o x i m a t e l y 14 days, and similar m e a s ur e m ent s were made f r o m J a n u a r y
to S ep temb er , 1980.
Shoots for growth analysis were decalcified in 10% h y d r o c h l o r i c acid,
and epiphytes scraped off. S h o o t material above the p u n c h e d hole in the
sheath was washed in sea water and divided into 'fresh growth', consisting
o f new leaves and the part of t he older leaves below the p u n c h e d hole, and
the remainder o f the shoot, f r om the p u n c h e d hole to the leaf tip. Both
c o m p o n e n t s were measured and dried to c o n s t a n t weight (70°C, 48 h).
Standing crops were calculated by summing t ot al leaf weight, and were
expressed per unit area. Leaf p r o d u c t i o n was calculated by dividing the
a m o u n t of new gr ow t h by the n u m b e r o f days elapsing b e t w e e n p u n c h i n g
and harvesting, and was expressed pe r unit area (Zieman, 1974). T u r n o v e r
rates or c o m m u n i t y r e p l a c e m e n t rates were calculated by dividing standing
crop by p r o d u c t i o n rate. A measure of relative leaf growth rate was ob-
tained by dividing p r o d u c t i o n by leaf standing crop (West and L a r k u m ,
1979), and expressing dr y weight as mg g-1 d a y - ] .
358

E p i p h y t e load on seagrass leaves

Two leaf blades were taken from each quadrat at the time of productivity
measurements, rinsed in seawater and frozen at P 1 0 ° C . Most of the epi-
phytes flaked off the frozen shoots easily, and the remainder were gently
scraped off. Shoots were then dried as described above. Epiphytes were
washed o n t o glass fibre papers (Whatman GF/C). Some samples were dried
to constant weight at 105°C for dry weight determination. Chlorophyll
extractions were carried out on other samples; the filters plus epiphytes
were ground, extracted in 90% aqueous acetone and optical densities of
extracts determined by the m e t h o d s described in APHA (1976) for peri-
p h y t o n . Chlorophyll a and p h a e o p h y t o n were expressed as mg m -2 of
seagrass leaf.
Complications arose when larger epiphytes were present, and an arbitrary
decision was made to include that part of the e p i p h y t e which shaded the
leaf under reproducible conditions. Each shoot was placed under a sheet of
plastic, and the outline of shoot and macroepiphytes traced. Macroepi-
p h y t e s were removed with forceps,, and their chlorophyll contents deter-
mined as described above. The tracings were p h o t o c o p i e d , and areas meas-
ured with a digitizer. The total projected area of each epiphyte was meas-
ured, along with t h a t part of it which covered the seagrass leaf; the ratio
between the two was calculated and used to apportion the total a m o u n t
of macroepiphyte chlorophyll. The calculated a m o u n t of chlorophyll was
expressed per unit leaf area and was added to the microepiphyte chlorophyll
load.
On one occasion, the seagrass in a quadrat was m a r k e d as described above,
leaf material harvested after 35 days and epiphyte loads on the new growth
determined. Ten shoots from each quadrat were rinsed in sea water, meas-
ured and divided into new growth and older leaf material; epiphytes were
removed for dry weight and chlorophyll analysed as described above.

E n v i r o n men ta l data

Water samples were collected at approximately m o n t h l y intervals at the

two sites, and analysed for nitrate+nitrite, a m m o n i u m , phosphate and
chlorophyll a using standard methods. Water sampling was also being carried
o u t on a regular m o n t h l y basis at the sites shown in Cambridge et al. (1986),
and data on temperature and light a t t e n u a t i o n were provided for those
sites closest to the seagrass sampling stations referred to here. Unpublished
incident radiation data for 1980 were provided by the D e p a r t m e n t of
Conservation and Environment, and the water depth and a t t e n u a t i o n coef-
ficient used to estimate total" daily radiation at the seagrass beds. Daily wind
data were from the Fremantle Port A u t h o r i t y . The total n u m b e r of hours
of wind at each range of speeds (10--20, 20--30, 30--40, > 4 0 knots) was
calculated for each day, and multiplied by the mid-point of the appropriate
 359

range. A mean dally estimate (knot h day -1 ) was calculated for each day
from leaf punching to harvest.
Each measured environmental parameter was p l o t t e d against time, and
the value at the mid-point between day of leaf punching and harvest was
used to calculate correlation coefficients between leaf production and
environmental parameters, using the SPSS programmes of Nie et al. (1975).

Periphyton on artificial substrates

Samplers were constructed of plastic (polyvinyl chloride; PVC) to each

hold 6 glass microscope slides. At each site, two samplers were attached by
rope to a sub-surface buoy, one with slides orientated vertically, the other
horizontally. Samplers were replaced each 2--3 weeks. There was some loss
of slides through storm damage. Harvested slides were each placed into
vials with filtered sea water and returned to the laboratory.
Before p e r i p h y t o n removal, light transmission was measured by placing
each slide on a stand made of vertical black PVC tubing containing a light
sensor (Li-Cor L a m b d a L1-185, Q u a n t u m Radiometer), which was clamped
so t h a t the sensor faced the end of the tube. To reduce optical problems
and prevent p e r i p h y t o n dehydration, the stand was submerged in an aquar-
ium filled with seawater. The light source above the p e r i p h y t o n slide was
a single beam from a fibre optics instrument (Schott-Mainz KL 150B;
150 watt, 15V Philips bulb). Light readings were taken at 400, 700 and
1,500 pE m -2 s-1 , using a blank reference slide, and the percentage reduc-
tion in light measured when a slide carrying p e r i p h y t o n was substituted.
Periphyton was then removed from each slide and washed o n t o filter
papers for dry weight or chlorophyll determination as described above.
Quadrats of plastic seagrass (0.1 m 2) were assembled using a base of
plastic-coated, woven steel mesh (25-mm squares). Strips of plastic approx-
imating the dimensions of a typical seagrass r a m e t or shoot with blades 6,
30 and 38.5 cm long and 11 m m wide, were attached to the grid with
staples and plastic-coated wire. The quadrats were anchored with steel pegs,
two quadrats per site.
Ten 'shoots' were harvested after 35 days, and returned to the laboratory
in sea water. They were cut into sections a b o u t 7.5 m m long, and two were
placed lengthwise on microscope slides for light transmission measurements
as described above. Periphyton was then scraped o f f for dry weight and
chlorophyll analysis.

Oxygen production

Shoots of Posidonia australis or plastic 'seagrass' were transported to the

laboratory at 10°C and maintained in an aerated tank of seawater at 20°C.
Oxygen evolution by leaf segments was measured using a Clark-type elec-
trode (Rank Bros., Bottisham, England) with an 8-ml vessel. Calibration was
360

with air-saturated sea water at the experimental temperature, 20°C, and with
a zero obtained after the addition of sodium dethionite.
Before each run, the cell was filledwith filteredsea water which had been
largely deoxygenated by bubbling through nitrogen containing carbon di-
oxide (295 vpm). Leaf segments were held upright by a plastic-coated wire
clip. The light source was as described for measuring light transmission
through periphyton, directed at right angles to the leaf surface through the
transparent walls of the vessel, so that the whole leaf surface was illumi-
nated. The amount of light reaching the leaf at each setting of the lamp was
estimated by substituting a half-vesselcontructed of perspex with a light sen-
sor behind it, for the experimental vessel so that refraction due to water and
perspex would be taken into account.
Segments of seagrass, ca. 2.5 c m long, were cut and placed into the cell,
and dissolved oxygen concentration recorded for 20 min after a lag phase
(see below). Epiphytes were then removed for chlorophyll measurement and
the process repeated with the leaf segments, n o w without epiphytes. Each
leaf segment was measured, ground and its chlorophyll a content measured
after extraction in 9 0 % aqueous acetone. Investigations were also carried out
with plastic seagrass segments carrying different epiphyte loads.

RESULTS

Shoot density, standing crop and leaf production

The mean leaf standing crop at Woodman Point, near the sewage works
outfall, was consistently less (by 65%) than at Carnac Island, the m o r e
oceanic site (Table I). The difference is partly explained by shoot densities,

TABLE I

O b s e r v a t i o n s o n s t a n d i n g c r o p s a n d p r o d u c t i v i t y o f Posidonia australis a t t h e t w o s t u d y
sites. All w e i g h t m e a s u r e m e n t s are in d r y w e i g h t , a n d m e a n s a r e a c c o m p a n i e d b y s t a n d a r d
errors

Measured property Carnac Island W o o d m a n Point

L e a f s t a n d i n g c r o p (g m -2 ) 215 -+ 32 95 -+ 23
S h o o t d e n s i t y ( n o . m -2 ) 522 * 32 368 +- 11
F l o w e r s ~ ( n o . m -2 ) 225 12
(g m -2 ) 286 16
L e a f p r o d u c t i o n ( g m -2 d a y -~ ) 2.7 ± 0.3 1.4 +- 0.3
Leaf turnover (day) 75 65
G r o w t h p e r s h o o t (rag d a y -~ ) 5.1 _+ 0.8 3.4 -+ 0.7
G r o w t h r a t e (rag g-~ d a y -] ) 12.9 -+ 1.2 15.4 -+ 1.3
Y e a r l y leaf p r o d u c t i o n ( g m -2 ) 949 475
L e a f + f l o w e r ~ p r o d u c t i o n ( g m -2 ) 1235 490
( L e a f + f l o w e r ) ~ p e r s h o o t (g) 2.37 1.32

Mean for August and September, including young fruits and embryos.
 361

which were on average 29% lower at W o o d m a n Point than at Carnac Island.

There were no distinct seasonal trends. Inflorescences were observed in
August and September at both sites, with 220--230 flowers m -2 on the two
occasions at Carnac Island, but only 10--14 at W o o d m a n Point.
In contrast to standing crop, leaf p r o d u c t i o n showed a distinct seasonal
pattern at both sites (Fig. 2), with high production in the summer m o n t h s ,
and low levels in winter and spring. The fall in p r o d u c t i o n in February--April
was more rapid at W o o d m a n Point. Mean leaf p r o d u c t i o n and growth per
shoot were significantly lower at W o o d m a n Point (50 and 33%, respectively)
than at Carnac Island.

CARNAC ISLAND
O---- WOODMAN POINT

~" 4,

\ /

J F M A M J J A S O N D J

Month

Fig. 2. L e a f p r o d u c t i o n o f P o s i d o n i a australis at t h e t w o sites s h o w n in Fig. 1. Vertical

bars are s t a n d a r d errors.

Shoots at Carnac Island generally consisted of 3--4 blades (0.37 g dry wt.
shoot-I), whereas at W o o d m a n Point the older blades tended to senesce and
break off earlier under a heavy e p i p h y t e load, and there were rarely more
than two blades per shoot (0.2 g dry wt. shoot-I). New growth, therefore,
made up a higher proportion of the shoots at W o o d m a n Point, and at Carnac
Island the total leaf biomass included a higher proportion of senescent mate-
rial. As a result of this, leaf replacement times were shorter at W o o d m a n
Point than at Carnac Island, while relative leaf growth rates (calculated as
increase in dry weight divided by total standing crop) were n o t significantly
different at the two sites.
The a m o u n t of leaf dry weight p r o d u c e d annually was calculated, and the
dry weight of a single crop of flowers and fruits was added to this; more than
twice as much production occurred at Carnac Island when compared with
362

W o o d m a n Point, and t he difference, although reduced, was still seen w hen

results were expressed on a 'per s h o o t ' basis (Table I). In subsequent calcula-
tions, th e leaf p r o d u c t i o n by the seagrass m e a d o w , calculated f r o m leaf den-
sity and average g r ow t h rate per leaf has been used as t he basis for compari-
son.

Environmental data

T h e mean c h l o r o p h y l l level in the water was significantly higher (P <

0.01; t test) at W o o d m a n Point than at Carnac Island, and so t o o was t he
c o n c e n t r a t i o n o f p h o s p h a t e (Table II). However, c o n c e n t r a t i o n s o f the in-
organic nitrogen c o m p o n e n t s were low, and n o t significantly d i f f e r e n t be-
t w e en th e sites. Ratios b e t w e e n nitrogen and p h o s p h o r u s were m u c h lower
t h a n th e ratio o f a b o u t 1 0 : 1 - - 2 0 : 1 , required to sustain high p h y t o p l a n k t o n
g ro w th rates, suggesting possible nitrogen limitation in t he water c o l u m n
( R y t h e r and Dunstan, 1971; G o l d m a n , 1976). As p o i n t e d o u t elsewhere
(Chiffings and McComb, 1981), low levels o f inorganic nitrogen are consis-
t e n t with rapid utilization o f available nitrogen in the Sound, while high
p h o s p h a t e c o n c e n t r a t i o n s , like high c h l o r o p h y l l levels, o c c u r in waters en-
riched b y e f f l u e n t f r o m i n d u s t r y and sewage t r e a t m e n t .

TABLE II
Properties of the water column at the two study sites. Data are means (with standard
errors) for February--September 1980 (n = 8)
Parameter Carnac Island Woodman Point

Chlorophyll a (~g 1-1) 2.1 ± 0.8 4.0 ± 1.0

Phosphate (#g 1-1) 13.0 ± 4.7 23.7 ± 9.2
(Nitrate + nitrite)-N (~g 1- ~) 5.2 ± 1.8 6.0 +- 1.8
Ammonia-N 0~g 1-1) 9.1 ± 2.4 8.4 ± 1.7
N:P ratios' 5.6 ± 1.8 5.3 ± 2.6

' Calculation by atom for inorganic nutrients.

Water t e m p e r a t u r e was at a m a x i m u m o f 24°C in J a n u a r y , and a m i n i m u m

o f 15.5°C in J u l y and August; W o o d m a n Point had a slightly higher tempera-
t u r e th an Carnac Island (at most 1°C) b e t w e e n May and July. Irradiance reach-
ing th e seagrass beds was at a m a x i m u m o f 21 X 106 J m -2 d a y - ' in J a n u a r y ,
and a m i n i m u m o f 5 X 106 J m -2 day -1 in June. T h e levels b e t w e e n the ex-
perimental sites were n o t consistently different; on average, t he light
reaching th e m e a d o w at W o o d m a n Poi nt was 92.4% o f t h a t at Carnac Island,
which was 78.3% o f t h a t at the surface.
Correlations were sought bet w e e n leaf p r o d u c t i o n and the following
variables: c o n c e n t r a t i o n s o f nutrients and chlorophyll, wind, light and tem-
perature. Significant correlations were f o u n d only b e t w e e n p r o d u c t i o n and
 363

t h e i n t e r r e l a t e d variables o f light (r = 0 . 7 4 7 , P = 0 . 0 0 2 ) and t e m p e r a t u r e

(r = 0 . 7 3 6 , P = 0 . 0 0 2 ) .

Epiphytes and periphyton

E p i p h y t e s r a n g e d f r o m m i c r o s c o p i c f o r m s such as d i a t o m s , t o larger
p l a n t s o f w h i c h t h e m o s t c o m m o n g e n e r a w e r e Ectocarpus and Myrionema
( P h a e o p h y t a ) , Polysiphonia, Laurencia, Ceramium and Melobesia ( R h o d o -
p h y t a ) , Ulva and Enteromorpha ( C h l o r o p h y t a ) . With few e x c e p t i o n s , simi-
lar t a x a were e n c o u n t e r e d as e p i p h y t e s o n leaves o r as p e r i p h y t o n o n glass
and plastic substrates. Ulva and Myrionema w e r e n o t f o u n d o n t h e glass
slides, while Calothrix was o n l y f o u n d o n glass. Ulva, Enteromorpha and
Calothrix were f o u n d o n l y at W o o d m a n P o i n t . F o r a m i n i f e r a n s a n d h y d r o i d s
were f o u n d o n all t h r e e substrates, b u t t h e r e w e r e v e r y few h y d r o i d s at
Carnac Island.
E p i p h y t e loads, e x p r e s s e d as c h l o r o p h y l l p e r u n i t leaf area, are s h o w n in
Fig. 3. T h e r e is a seasonal t r e n d , w i t h low loads at t h e e n d o f s u m m e r , and
an increase in m i d - a u t u m n ; t h e loads were m u c h h i g h e r at W o o d m a n P o i n t ,
as was t h e a m p l i t u d e o f t h e g r o w t h curve. T h e r e was a m a r k e d increase in
e p i p h y t e load in mid-April at this site, and this c o r r e s p o n d e d w i t h the
s u d d e n fall in leaf p r o d u c t i o n at t h a t t i m e (cf. Fig. 2). E p i p h y t e d r y w e i g h t
s h o w e d similar t r e n d s t o c h l o r o p h y l l at Carnac Island, b u t w e r e m o r e erra-
tic at W o o d m a n P o i n t , w h e r e a high load in A u g u s t - - S e p t e m b e r is a t t r i b u t e d
126-

105-
/ \
/ \
/ \
E 84- / ~ CARNAC ISLAND
~ - W O O D M A N POINT
/
/ \
~ 63- / \
o
Z / \
/ \
~ 42-
/ \
/
21-
/ \,~/ \\ j

J F M A M J J A S O N D J

Month

Fig. 3. Epiphyte toad (as epiphyte chlorophyll expressed per unit area of seagrass leaf) at
Woodman Point, near to the sewage effluent, and Carnac Island, in more oceanic water
(Fig. 1).
364

to sand particles trapped by macroepiphytes and hydroids. On one occasion,

a comparison was made between mean e p i p h y t e loads on new growth and the
remainder o f the harvested leaves; on new growth at W o o d m a n Point chloro-
phyll epiphyte load was 0.28 (SE 0.02) pg cm-:, as compared with 0.12
(0.03) at Carnac Island; on the older leaves the figures were 6.57(1.7) at
Woodman Point, and 1.65(0.50) at Carnac Island (n = 5).
Rates o f p e r i p h y t o n biomass showed generally similar seasonal trends to
epiphyte biomass (Fig. 4), with higher loads at W o o d m a n Point. Changes
in p e r i p h y t o n dry weight were more erratic, again because of sand grain
accumulation.
1.8-
Z~ CARNAC ISLAND
O----- WOODMAN POINT

1.5-
/
A/
f / J
E
1.2- / \ /
/ /
\
~. 0.9-
o /
/
~ 0.6-

0.0 J F M A M J J A S O N D J

Month

Fig. 4. Periphyton load (as chlorophyll expressed per unit area on glass slides). Gaps in
the data were due to loss of samples. The two sites are as shown in Fig. 1.

For pooled data there was a high correlation between epiphyte and peri-
p h y t o n chlorophyll a levels, and between epiphyte and p e r i p h y t o n dry
weights (Table III). This provides strong circumstantial evidence that the
factors controlling p e r i p h y t o n growth and epiphyte load are the same.
Correlations were sought between e p i p h y t e or p e r i p h y t o n chlorophyll on
the one hand, and water column n u t r i e n t and chlorophyll concentrations,
wind, light and temperature on the other. The only significant correlations
were with water column chlorophyll concentrations; for the epiphytes, r =
0.811, P = 0.007; for the p e r i p h y t o n , r = 0.900, P = 0.019. This is con-
sistent with the suggestion t h a t the factors controlling p h y t o p l a n k t o n bio-
mass are similar to those which control the loads of p e r i p h y t o n and epi-
phytes.
 365

TABLE III

Correlations between epiphyte and periphyton loads

Periphyton chlorophyll a Periphyton dry weight

Correlation Significance Correlation Significance

coefficient r coefficient r

Epiphyte
chlorophyll a 0.835 0.003 0.683 0.020
Epiphyte
dry weight 0.883 0.010 0.830 0.020

Light reduction by epiphytes

T h e r e was a clear r e l a t i o n s h i p b e t w e e n light r e d u c t i o n a n d p e r i p h y t o n

load (Fig. 5), t h e d a t a a p p r o x i m a t i n g t o t h e f o l l o w i n g f u n c t i o n (n = 35;
r = 0.725;P<: 0.001):

f(x) = 100[1-e(-0.5x)]
The r e l a t i o n s h i p f o r d r y w e i g h t (n = 23; r = 0 . 7 3 1 ; P <: 0 . 0 0 1 ) was:
f(x) = 100[1-e(-°.2x)]

L i g h t r e d u c t i o n b y p e r i p h y t o n o n plastic seagrass gave a similar c u r v e t o

t h a t o f p e r i p h y t o n o n glass slides.
Using t h e curves f o r light r e d u c t i o n b y k n o w n p e r i p h y t o n loads, o n e m a y

l°°t J
J
80

60-

40-

20-

0
o.o 0'.2 11o l'.s 2'.0 2'.s 31o 3'.2 4'.0
PerJphylon C h l o r o p h y l l a /~g c m - 2

Fig. 5. The relationship between light reduction and periphyton chlorophyll on glass
slides. The slides were collected from the field, and light reduction, as proportion of in-
cident PAR, measured in the laboratory.
366

estimate the light reduction by epiphytes on seagrass leaves. The mean epi-
p h y t e chlorophyll load at Carnac Island was 0.62 gg cm -2, which represents
a mean light reduction of 31%. At Woodman Point the mean load was 5.41,
which converts to a light reduction of 96%. It m u s t be remembered, how-
ever, that epiphyte loads are relatively low on new leaf material. Using the
loads recorded above for this new growth, light reduction would be 7% at
Carnac Island, and 15% at Woodman Point. Using this figure for new growth,
and the above figures for the older leaf material, the overall reduction in
light was a b o u t 35% at Carnac Island, and 63% at W o o d m a n Point.

Oxygen production
The trapping of oxygen within leaf lacunae would lead to underestimates
o f p h o t o s y n t h e t i c rates by the m e t h o d used here. When an experiment was
started there was a lag phase, usually lasting 10 min, after which oxygen
production rate increased to a constant level; presumably the lag phase was
the time taken for the severed leaf lacunae to become saturated with oxygen.
The rate of p r o d u c t i o n of oxygen after the lag phase was used in calcula-
tions. There was no lag phase when measurements of photosynthesis were
made using p e r i p h y t o n on segments of plastic seagrass.
Figure 6 shows p h o t o s y n t h e t i c rates before and after e p i p h y t e removal,
plotted against light intensity. Both are typical photosynthesis versus irra-

200 ......
150- /
V
/
.E
E 100- /
?
i -I -I
I so- I I #,t
tI J'
0 /I
//
//
- 50 ~ I//I

- 100 680 12100 18=00 24=00 30=00 36v00

Par (/~E m -2 sec -1)

Fig. 6. Rates o f p h o t o s y n t h e s i s o f segments f r o m seagrass leaves, w i t h ( a b o v e ) and w i t h -

o u t (below) epiphytes, at different light intensities. Vertical bars are standard errors;
n=5.
 367

diance curves (e.g. Steemann Nielsen, 1975; Drew, 1979), with light satura-
tion at a b o u t 1500 p E m -2 s-2. Similar curves were obtained when rates were
calculated on the basis of seagrass chlorophyll weight, rather than dry
weight.
Rates of p h o t o s y n t h e s i s of seagrass segments which naturally lacked epi-
phytes were similar to those from which epiphytes had been scraped, sug-
gesting t h a t e p i p h y t e removal did not damage the leaves. Oxygen produc-
tion rates at light saturation were measured for segments of plastic seagrass
carrying k n o w n p e r i p h y t o n loads, and the relation between the two p l o t t e d
(Fig. 7); this gave a linear correlation (regression through the origin) of 0.88
(P < 0.05). K n o w i n g the a m o u n t of chlorophyll present in the epiphytes,
their oxygen p r o d u c t i o n at saturating light intensities could be estimated
from the graph.

/
360-

~ 300- • • - -

E 240-

.~ 180-

o~
c 120-

×
0

60-

Chlorophyll a (/J.g c m -2)

Fig. 7. The relationship between oxygen production at saturating light and periphyton
chlorophyll a load, both expressed per unit area.

For example, on one occasion the difference between o x y g e n p r o d u c t i o n

by a seagrass leaf segment before and after epiphyte removal, was 235.2 pg
cm -2 min -1 at saturating light. The epiphyte load on the seagrass leaf was
3.74 pg cm -2, which according to Fig. 7 might be expected to produce oxy-
gen at a rate o f 236 + 26 (SD) pg cm -2 min -1. This suggests t h a t the photo-
synthetic performance o f the p e r i p h y t o n is similar to t h a t o f the epiphytes.

DISCUSSION

A recurring trend in these results is t h a t the seagrasses at Carnac Island are

more vigorous than those at W o o d m a n Point, where a 33% lower growth per
368

s h o o t and a 29% less dense m e a d o w resulted in 50% less leaf production.

There is evidence from aerial p h o t o g r a p h y that the seagrass m e a d o w s at
W o o d m a n Point were once denser than t h e y are now, and it is possible that
t h e y once resembled more closely those at Carnac Island. There are higher
epiphyte loads at W o o d m a n Point, and higher chlorophyll and phosphate
levels in the water column there. While these observations are consistent
with the suggestion that increased epiphyte loads are associated with increas-
ed eutrophication in Cockburn Sound (Cambridge et al., 1986), and that
nitrogen is more critical to algal growth than is phosphorus in this system
(Chiffings and McComb, 1981), t h e y do n o t in themselves provide evidence
for a causal connection b e t w e e n eutrophication, increased epiphyte growth
and reduced seagrass production.
Leaf p r o d u c t i o n rate was significantly correlated with light and tempera-
ture, which are intercorrelated, so that their separate effects are difficult to
disentangle from such analysis. However, the laboratory studies show that
light intensities in the Sound at the level of the seagrass m e a d o w s would be
barely sufficient to light-saturate seagrass photosynthesis, especially when it
is born in mind that illumination in the laboratory was at right angles to the
leaf surface; that there is m u c h mutual shading in a seagrass m e a d o w ; that
there are non-photosynthesising tissues to be supported; and that there are
periods of darkness and low light intensities in the field. Even in these
shallow waters, light must be of critical importance and any substantial in-
crease in shading will presumably affect the growth of the seagrasses.
Cambridge et al. (1985) have already suggested that shading by phyto-
plankton was n o t responsible for the widespread loss of seagrass from
Cockburn Sound. The a m o u n t of light reaching the seagrass m e a d o w s at the
t w o shallow (3 m) sites studied here did not differ significantly, and differ-
ences in seagrass p r o d u c t i o n were n o t correlated with chlorophyll levels in
the water.
The ability o f 'epiphytes' to colonise artificial substrates as well as living
seagrass, the high correlations between the biomass of the t w o and their
similar p h o t o s y n t h e t i c performance are consistent with observations else-
where that submerged plants act largely as neutral substrates for epiphyte
growth (Catteneo and Kalff, 1979; B o r u m and Wium-Andersen, 1980), al-
though transfer o f materials can occur b e t w e e n host and epiphytes, and m a y
be especially i m p o r t a n t at low ambient nutrient levels (for reviews and other
references see Harlin, 1980; Bulthuis and Woelkerling, 1983; Orth and Van
Montfrans, 1984). The enhanced growth of epiphytes on deteriorating
meadows in Cockburn Sound, and on seedlings transplanted into them
(Cambridge et al., 1986) cannot, therefore, be reasonably ascribed to leakage
of nutrients from senescing seagrass leaves. The present w o r k shows that
periphyton (and epiphytes) grow more rapidly and build up larger standing
stocks in nutrient-enriched waters, and this is presumably, like phyto-
plankton growth, a response to nutrient enrichment. A further complexity
is the possibility of reduced grazing of epiphytes in nutrient-rich water
 369

(Orth and Van Montfrans, 1984) which we have not investigated quantita-
tively. However, the periphyton on glass slides was not affected to any ex-
tent by grazing, and the few slides which showed gastropod tracks were
discarded.
The shading of leaves by epiphytes is clearly significant, and is higher at
Woodman Point (63% overall) than at Carnac Island (35%). F r o m Fig. 5, this
would bring about a decrease of 80% at PAR 500 ~E m -2 s-1 at W o o d m a n
Point, while at Carnac Island the reduction would be 35%. These results can
be compared with those of Sand-Jensen (1977), who estimated the reduc-
tion in photosynthesis of Zostera marina L. by epiphytes to be 31%. The
results are also consistent with the observations of Bulthuis and Woelkerling
(1983), who have shown that epiphyte growth markedly reduces the time
during which positive photosynthesis is possible for developing leaves of
Heterozostera tasmanica (Martens ex Aschers.) den Hartog in Western Port
and Port Phillip Bays, Victoria.
When leaf production observed in the field was graphed against observed
epiphyte load (Fig. 8), the resulting curve showed a negative log-linear cor-
relation (r = 0.87; P < 0.001), and a similar curve was obtained when leaf
standing crop was plotted against e p i p h y t e load. The relationship m a y be at-
tributed to that between epiphyte loads and their attributed light reduction.
Indeed the relation between leaf p r o d u c t i o n and light reduction due to epi-
phytes (Fig. 8) gave a correlation coefficient of - 0 . 9 0 (P < 0.01), and with
a m o u n t of periphyton growing on glass slides gave a correlation o f - 0 . 7 9
(P < 0.01). It is clear that high e p i p h y t e loads are correlated with low leaf
production.
3.6-

3.0- I)0

"o

~.
2.4-

1.8-
!
Z
: t+2.

0.6-

0.0
~0 ,5 ~0 8'0 t~0 t~0 t~0 ~o ,~ io io t~o
Epiphyte Chlorophyll a (mg Ill- 2 Lear-~) % Light Reduction by Eplphytes

Fig. 8. The relation b e t w e e n seagrass leaf p r o d u c t i o n m e a s u r e d in the field and e p i p h y t e

load; left, load expressed as e p i p h y t e c h l o r o p h y l l per unit leaf area; right, using light re-
d u c t i o n brought a b o u t by d i f f e r e n t e p i p h y t e loads as e s t i m a t e d f r o m the relationship in
Fig+ 5. Data bulked f r o m b o t h sites over the period J a n u a r y - - O c t o b e r .
370

The calculations reported in this paper necessarily contain a number of

approximations, but there seems little doubt that shading by epiphytes has
a marked effect on the productivity of seagrasses, sufficient to account for
the reduced growth of seagrasses in nutrient-enriched waters, and providing
support for the suggestion (Cambridge et al., 1986) that enhanced epiphyte
growth following nutrient enrichment has been responsible for seagrass de-
cline in Cockburn Sound.

ACKNOWLEDGEMENT

We are indebted to the Department of Conservation and Environment,

Western Australia, for financial assistance and continuing interest.

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