Aileen T Shau-Hwai

M A Munira

M Adlim

Muhd Nasir

Zulfigar Yasin

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OPEN ACCESSAquaculture, Aquarium, Conservation & Legislation
International Journal of the Bioflux Society Research Article
Volume 5 | Issue 1 Page 40 
AACL Bioflux
http://www.bioflux.com.ro/aacl
Determination of seasonal skeletal growth layers 
in Porites lutea colonies from Teluk Nyior reefs, 
Langkawi Island, Malaysia
1Muhd. Nasir, 2Zulfigar Yasin, 2Aileen T. Shau-Hwai, 3M. Adlim, 4M. A. Munira
1Departement of Biology, Syiah Kuala University, Banda Aceh, Indonesia; 2School of Biological Sciences, Universiti Sains Malaysia, 
Minden, Penang, Malaysia;  3Department of Chemistry, Syiah Kuala University, Banda Aceh, Indonesia; 4Politeknik Kesehatan Aceh, 
Banda Aceh, Indonesia.
Introduction 
Long-lived massive corals can contribute to the monitoring of 
coral reef environment through measurable growth records in 
the annual growth layer thickness. Annual growth layer pattern 
in massive coral skeletons provide means to “retrospectively 
monitor”. These can provide a historical perspective, which is 
able to determine environmental changes and can help to es-
tablish limits of coral growth. Furthermore, records from cor-
als may provide information about long-term variability in the 
performance of coral reefs, allowing unnatural changes to be 
distinguished from natural variability.
The studies of skeletal growth in massive coral have been in-
tensively reported (Buddemeier & Kinzie 1975; Barnes et al 
2003; Carricart-Ganivet 2004; Gourbert et al 2001; Lough 
& Cooper 2011). Coral growth determinations using alizarin 
staining method have been investigated by Lamberts (1978). 
Technically, the annual increments of growth of shallow-wa-
ter of Porites lutea were calculated using seasonal fluorescent 
banding (revealed by UV-light) and alizarin staining. The tim-
ing and seasonality of skeleton construction has been calibrated 
using alizarin marked coral (Mizrachi et al 2010). There have 
been written many publications employing alizarin for coral 
growth staining. However, there are scarce reports on combi-
nation of the alizarin staining method with UV artificial lumi-
nescence for determination of coral seasonal layers particularly 
on Porites lutea. Hence, the objective of the present study was 
to investigate the coral growth layer and also intends to ascer-
tain seasonal layers in P. lutea skeletal using alizarin staining. 
This paper is contributing the advance techniques to study the 
annual coral growth pattern of massive corals.
Material and methods
Study area
The study was carried out at Teluk Nyior reef flat, Langkawi 
Island, Malaysia. The island is situated in Strait of Malacca, 
at latitude 6o 26’ N; longitude 99o 45’ E on the west coast of 
Peninsular Malaysia (Figure 1) and the detail map of the sam-
pling location was presented in Figure 2. The site is approxi-
mately 40 km from the main town of Kuah, Langkawi Island. 
The reef slope, where the studies have been made is visibly flat.
 
Alizarin staining
Samples of P. lutea with size 20-30 cm diameter were collected 
from study sites seasonally. Approximately 22 colonies were 
stained with 10-ppm alizarin red for 24 hours (Lamberts 1978; 
Harriot 1999) during for both wet (17 October 2001) and dry 
season (17 January 2002). Staining Porites colonies during wet 
and dry season were done to recognize characteristic of growth 
layers. After staining the corals were placed on their original 
location and allowed to grow until completely one year. 
Abstract. This study includes seasonal layer study of Porites lutea using combination of alizarin staining and UV illumination methods and the 
correlation with climate data. The growth layers were recognized by using alizarin staining method, which was marked on live tissue of corals 
P. lutea. Under the UV-light, dark fluorescent layers represented the wet season (June to November) and light fluorescent layers corresponded 
to dry season (December to May). The results revealed that the alizarins integrated into newly forming skeleton which looks as pink color. The 
color remains as a permanent implant indicating the place of calcification during the experiment. Seasonal layers (wet and dry season) can be 
used to determine the skeletal growth thickness yearly, since the season cycles are relative regular in whole year. Recognizing and ascertaining 
the dark and light fluorescent layers were useful for retrospectives analysis of coral age prediction.
Key Words: alizarin, fluorescent, coral growth, Porites lutea, season.
Copyright: This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted 
use, distribution, and reproduction in any medium, provided the original author and source are credited.
Corresponding Author: M. Nasir, muhd nasir@fmipa.unsyiah.ac.id
Nasir et al 2012
Volume 5 | Issue 1 Page 41 
AACL Bioflux
http://www.bioflux.com.ro/aacl
Coral sample cutting 
One year after staining, the samples were collected and sent to 
the laboratory. The samples were cut vertically using an elec-
tric saw (Model L80P, Light outer, S. D. K. Co. Ltd) in a dark 
room to avoid light oxidation reaction on organic material in the 
coral skeletal. At least 2 cm thick slices were removed from the 
center of the colony (Figure 3).  The corals slices were washed 
by distilled water and then air-dried. UV artificial luminescent 
was used to observe seasonal growth layers pattern using UV-
lamps (4 black tube x 20 watt) (Figure 4).
Results and Discussion
Seasonal layers
Fifty-two colonies of Porites lutea have been alizarin stained, 
twenty-two colonies of which were stained during wet season 
and the thirty colonies were stained during dry season. The sam-
ples were placed in Teluk Nyior reef for one-year growing from 
the date of staining, then after one year the samples were rec-
ollected.  However, only thirty-six colonies were taken, which 
were thirteen colonies of wet season treatment and twenty-three 
colonies of dry season samples. Sixteen colonies have been lost 
in the sea, there were moved by sea current. Annual growth lay-
ers of P. lutea have been observed in skeletal slices. The growth 
layers were recognized by using Alizarin staining method, which 
was marked on live tissue of corals P. lutea (Lamberts 1978; 
Mizrachi et al 2010; Swart et al 2002). The alizarin was used 
to definite seasonal layer in P. lutea skeletal.  
Alizarin in P. lutea skeletal was marker permanently, which can 
be directly observed in skeletal slices of P. lutea samples and 
photographed. One year after staining, the pink line of alizarin 
was clearly observable in P. lutea skeletal. Under the UV-light, 
dark and light fluorescent layers can be differentiated. Alizarin 
line stained on coral colonies during wet season (17 October 
2001) appears in dark fluorescent layers as shown in Figure 5. 
Unlike in wet season treatment sample, alizarin line stained 
during dry season (17 January 2002) appears in light fluores-
cent layers as shown in Figure 6. These results indicated that 
dark layers in P. lutea skeletal from Teluk Nyior reef were de-
posited during wet season (June to November) and light layers 
form during the dry season (December to May). 
It is interesting to note that the alizarin stain stained in wet sea-
son exhibited the line mark on the dark area (under UV-light) 
in coral skeletal as shown in Figure 5. This line marks are con-
sistent in other samples. Seventy percent of the samples in this 
treatment show a consistent position of alizarin line mark, which 
was on dark area. Unlike wet seasonal samples, the alizarin mark 
Figure 1. Map of Pulau Langkawi, showing the Teluk Nyior study 
area.
Figure 2. Description of place where Porites colonies were 
collected on  Teluk Nyior reef, Langkawi Island. 
Figure 4. Illustration of mechanism to determine seasonal  Layers 
using UV-light. 
Figure 3. Illustration of a Porites colony sliced vertically.
Nasir et al 2012
Volume 5 | Issue 1 Page 42 
AACL Bioflux
http://www.bioflux.com.ro/aacl
of coral stained in dried season however appeared on the light 
area (under UV light) of coral skeletal (Figure 6). This is also 
consistent on the other dried seasonal treatment samples. About 
eighty percent of samples consistently showed the alizarin mark 
on the light area as shown in Figure 6.  This finding is consist-
ent with previous report employing X-ray diffraction to observe 
the skeletal layers. As state by Scoffin et al (1989) that the dark 
area of florescence bands were formed on the wet season. 
Living corals P. lutea were subjected to alizarin stain, which 
is dissolved in seawater appear as purple color. The alizarins 
integrated into newly forming skeletons, which look as pink 
color. The color remains as a permanent implant indicating the 
zone of calcification during the experiment. The pink line in P. 
lutea skeletal samples was varying in evident, 70% of samples 
colonies were bright clear and the rest samples have blur pink 
line in their skeletal. As Lamberts (1978) suggested that alizarin 
coloration is influenced by the total amount of living tissue de-
positing the dye, the concentration of the dye in solution, length 
of exposure to the dye, and biological activeness of the calcify-
ing tissue. Dark layer in P. lutea skeletal might be correlated to 
the absorption of humic acids produced by plants and flowed 
by fresh water run-off to the sea as reported by Isdale (1984). 
Having identified in this study that the dark area represents coral 
growth in the wet season and the light area is in dry season, it 
is possible to predict the age of corals on the basis of dark and 
light layer. Since the season cycles are relative regular in whole 
year. Several authors including Buddemeier & Kinzie (1975), 
Barnes & Lough (1993), Hudson et al (1994) and Priess et al 
(2002) have introduced these predictions. Furthermore, Wolanski 
et al (2004) predicted changes in salinity, nutrient, pH, incident 
light, temperature or various changes in the chemical constitu-
ents of seawater on the basis of seasonal layer. 
Recognizing and ascertaining the dark and light fluorescent layers 
were useful for retrospectives analysis of coral age predicting. 
In addition, dark and light fluorescent layers in P. lutea skeletal 
from Teluk Nyior reef may be the effect by terrestrial runoff. 
These layers also have the potential to provide information of 
regional variations in rainfall, river flow and changes in land. 
They also provide a means to assess the risk to corals, coral 
communities and corals reefs from runoff and changes in land-
use, pollution, oceanography, and coastal and reef management 
(Al-Rousan et al 2007; Ahmad et al 2011; Schiedek et al 2007). 
Conclusions
The alizarins integrated into newly forming skeletons, which 
look as pink color. The color remains as a permanent implant 
indicating the calcification place during the experiment. The 
pink line in P. lutea skeletal samples was varying in evident, 
70% of samples colonies were bright clear and the rest samples 
have blur pink line in their skeletal. The alizarin stained line can 
be used to determine the skeletal growth thickness, to ascertain 
seasonal layers (wet and dry season). Dark fluorescent layers 
represent coral growth in wet season and light fluorescent lay-
ers correspond to the growth in dry season. 
Acknowledgement 
Appreciation is dedicated to Universiti Sains Malaysia Reef 
Research Group and The Technical Assistants at Muka Head 
Marine Research Station for their assistance and logistical support.
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H., 2007. Heavy metal contents in growth bands of Porites cor-
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Figure 5. The Porites skeletal slice showing alizarin-stained line 
on wet season (a). The stained line was located in dark area of 
fluorescent layers; (b) Light fluorescent layer, (c) Dark fluorescent 
layers. 
Figure 6. The Porites skeletal slice showing alizarin-stained line 
on dry season. (a). The stained line was located in dark area of 
fluorescent layers; (b) Light fluorescent layer, (c) Dark fluorescent 
layers.



Determination of seasonal skeletal growth layers in Porites lutea colonies from Teluk Nyior reefs, Langkawi Island, Malaysia

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Determination of seasonal skeletal growth layers in Porites lutea colonies from Teluk Nyior reefs, Langkawi Island, Malaysia

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