Epinephelus suillus
I. Introduction
At present, approximately one-third of CO2 has already been absorbed by the earths
atmosphere that has accumulated for the past 100 years. This has led to the increase in the partial
pressure of carbon dioxide in the oceans and decrease in pH levels (Caldeira & Wickett 2003,
Royal Society, 2005, and German Advisory Council on Global Change, 2006). Moreover, this
level has been the highest CO2 concentration than any point in the past 800,000 years (Lthi D,
et al. 2008) and has the fastest rate experienced in the last 300 million years (Hnisch B, et al.,
2012).
It is known that ocean acidification affects a wide diversity of marine organisms. Most of
the research conducted on CO2-driven ocean acidification concerns on corals that focuses on
calcification and effects on other calcifying organisms. However, studies on the impact on fishes
are more limited both in understanding and in a variety of species. The early life stages of other
marine organisms particularly on fishes are also likely to be sensitive to these changes for they
are essential for replenishment and connectivity (Portner et al. 2004). Increasing attention has
been given to the impacts of ocean acidification on the early development stages of marine
species, but experiments in relation to these scenarios have provided mixed results (Munday, et
al., 2011 and Nilsson GE, et al., 2012). The early development stage is a vital phase in marine
fishes as this is the life stage which is more susceptible to mortality (Houde E., 1997). Therefore,
it is important to study the effects of ocean acidification on the early life stages of fish in
abundant supply compared to the other species in the genera (Kohno, 1987). In addition, it is one
of the most widely distributed tropical species with significant ecological and economic value.
Its life history and morphology are similar with other high-value, tropical fishes and thus their
use in these experiments provides a useful perspective of possible ocean acidification impacts on
B. Objectives
This study aims to examine the development of the eggs and larvae of E. suillus, and their
rate of growth and survival when subjected to elevated CO2 levels (380, 600, 750 and 1000
ppm).
C. Hypothesis
CO2-driven ocean acidification will have damaging effects on the morphology of the
atmosphere at 278 ppm has risen to approximately 390 ppm in 2010 which causes significant
When CO2 is absorbed in seawater, it reacts with H2O to form H2CO3 (carbonic acid).
In turn, the carbonic acid will break into H+ ions and HCO3- ions (bicarbonate ions). Carbonic
acid will break down into H ions and HCO3- (bicarbonate ions). Increase in H+ ions will lower
the pH thereby increases the acidity. The CO32- (carbonate ions) react with these H+ ions to
form more bicarbonate ions. Carbonate ion concentrations decreases while H+, H2CO3 and
HCO3- concentrations increase. When carbonate ions are less available, calcium carbonate
(CaCO) dissolution is more likely to occur, and consequently its formation is less likely to occur
(ASOC, 2010).
B. Impacts on vertebrates
The impacts of ocean acidification affects all marine life. However, studies on the effects
on vertebrates has been very limited but there are indication of detrimental effects on fishes
although these impacts are still unknown. It has been found out that the short-term effects of
ocean acidification on fishes includes acidbase status, respiration, blood circulation, and
nervous system functions, while long-term effects include reduced growth rate and reproduction
(Guinotte, JM and Fabry, VJ, 2008). Also, acidification of body fluids of marine animals
resulting from increasing hypercapnia occurs rapidly, in a matter of hours (Royal Society, 2005).
It is also found out that hypercapnia lowers respiratory rates and protein synthesis rates, and
The experiment will be conducted at the Marine Pollution and Ecotoxicology Laboratory
A. Egg Collection
E. suillus eggs will be supplied by the aquaculture department of SEAFDEC. Throughout
B. Experimental Design
A flow-through system will be used in the setup and will be equilibrated with different
CO2 concentrations. The aquaria will be placed in a water bath to minimize water temperature
fluctuations. The containers will be covered tightly with lids and a single hole will be cut for
seawater and gas tubing to fit inside. The seawater will be pre-bubbled with CO2 until it reaches
the current level of atmospheric CO2 which is approximately 380 ppm and three values projected
to occur in the future within the current century which is 600, 750 and 1000 ppm as
recommended by EPOCA protocol (Barry, JP, et al. 2010). There will be three replicates for
each treatment. CO2 concentration will be measured in ppm units using a CO2 gas probe. The
pH of the water will be monitored throughout the experiment using a pH meter. Total alkalinity
and dissolved organic carbon will be measured on subsamples and the carbonate system will be
Three hundred eggs will be randomly assigned to each treatment (100 eggs for each
replicate). Five randomly selected eggs per aquaria will be monitored every fifteen minutes one
hour after the fertilization for the duration of five hours, and every three hours for the next 24
Dead cells will be removed from the treatments for morphometric analysis. E. suillus will
be photographed and measured for each developmental stage and will be compared to the
controlled. Each of them will be dried to a constant mass to obtain the dry mass.
C. Computations
Hatching rates will be determined using the formula:
.
=
.
=
= 100
D. Statistical Analyses
the four pCO2 treatments will be tested with one-way ANOVA. Tukeys HSD multiple
comparison test will be used to determine differences in pCO2 levels. Any bias in these data will
be explored by residual analysis and if detected the data were transformed as noted in the
ANOVA tables. A contingency table will be used to test whether the number of larvae settling
IV. Timeframe
J J A S O N D J F M A
Capsule proposal
Proposal first draft
Final proposal
Experiment proper
Thesis first draft
Thesis second draft
Final thesis
V. Budgetary requirements
Amount (Php)
Grouper eggs 100
Transportation 100
CO2 750
Others 1000
TOTAL 1950
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