The Effects of Clove Oil in Color Changes and ... · The Effects of Clove Oil in Color Changes and...

The Effects of Clove Oil in Color Changes and Zooxanthellae Density on Porites lobata

Sri Wahyuni Rahim *, M. Natsir Nessa, Dody D. Trijuno and M. Iqbal DjawadFaculty of Marine Science and Fisheries, Hasanuddin University, Makassar

Abstrak

Penangkapan merusak seperti penggunaan sianida dalam penangkapan ikan hias telah menimbulkan kerusakan terumbu karang sehingga disarankan penggunaan minyak cengkeh yang dianggap ramah lingkungan. Penelitian ini bertujuan untuk menganalisis perubahan warna, kepadatan zooxenthella dan indeks mitotik pada karang Porites lobata setelah pemaparan beberapa konsentrasi minyak cengkeh. Disain eksperimen adalah rancangan faktorial yang terdiri dari 6 variasi perlakuan konsentrasi dan 5 variasi perlakuan waktu dengan 3 kali pengulangan. Karang dimasukkan ke dalam akuarium berukuran 1.0x0.5x1.0 m yang berisi larutan minyak cengkeh yang berbeda konsentrasinya (kontrol, 20, 30, 40, 50, dan 60 ppm) selama 5 menit kemudian dipindahkan ke akuarium yang berisi air laut yang bersih selama 60 detik lalu dipindahkan ke bak penampungan untuk proses pemulihan. Pengamatan dilakukan setiap hari untuk perubahan warna karang sampai hari ke-17, sedangkan perubahan kepadatan zooxanthellae dan indeks mitotik dilakukan pada hari ke-1, 5, 9, 13 dan 17. Hasil penelitian menunjukkan bahwa tidak terjadi perubahan warna karang sampai hari ke-17 setelah pemaparan minyak cengkeh 20–60 ppm pada karang Porites lobata dan tidak ada perbedaan yang signifikan pada kepadatan zooxanthellae dan indeks mitotik karang Porites lobata terhadap variasi konsentrasi minyak cengkeh, tetapi terdapat perbedaan terhadap variasi waktu (hari). Hal ini menunjukkan bahwa pemaparan minyak cengkeh pada konsentrasi 20–60 ppm tidak mempengaruhi perubahan warna karang, kepadatan zooxanthellae dan indek mitotik karang Porites lobata.Kata Kunci: minyak cengkeh, zooxanthellae, Porites lobata

Abstract

Destructive fishing such as using cyanide in capture of ornamental fish has caused damage to the coral reefs, this is suggested the use of clove oil which considered eco-friendly fishing. This study aims to analyze the changes of color, zooxanthellae density and mitotic index of coral Porites lobata after exposure to several concentrations of clove oil. Experimental design was a factorial design, which consists of six concentrations and five time treatments with 3 replication. Coral was placed into the 1.0x0.5x1.0 m aquarium that contains clove oil solution in different concentrations (control, 20, 30, 40, 50, and 60 ppm) for 5 min and then transferred to a tank containing clean seawater for 60 seconds and then removed to tanks for the recovery process. Observation was made everyday for coral color changes in 17 days period and on day 1, 5, 9, 13 and 17 for zooxanthellae density and mitotic index. The results showed that the were no changes in Porites lobata colors until the 17th day after exposure to 20–60 ppm of clove oil and there was no significant difference in the zooxanthellae density and mitotic index of the Porites lobata on concentration variation, while zooxanthellae densities and mitotic index showed significant difference on time (day) variation. This suggested that exposure to clove oil at a concentration of 20–60 ppm did not affect the color change, zooxanthellae density and mitotic index of Porites lobata.Keywords: clove oil, zooxanthellae, Porites lobata

Journal of Indonesia Coral Reefs 1(3) (2012) 205-216Printed ISSN : 2089-8231

*Corresponding Author: [email protected]

206


INTRODUCTIONDestructive fishing such as the use of

cyanide in capture of ornamental fish has caused damage to coral reefs. This practice change the structure of the habitat and affect diversity, composition, biomass and production of the organisms associated with coral reef habitats. Jones and Steven (1997) have reported that sodium cyanide (NaCN) at high dose (2x10-1 M) will easily kill coral; moderate dose (2x10-2 M) corals will lose zooxanthellae (symbiotic algae) known as the bleaching process and the low dose (2x10-3 M/2x10-4 N) of cyanide causes loss zooxanthellae but not until bleaching or discolouration phase. The average of coral respiratory rate was also reduced to 10–90% because of the cyanide poison. Fish that are exposed to cyanide solution are rapidly asphyxiated, eventhough they are transferred to clean seawater, captured fish suffer a suite of physiological disruptions, and as many as 80% die as a consequence (Rubec et al., 2001).

Most serious aspects of cyanide fishing impact on coral reefs. Subandi (2004) exposed to cyanide concentration of 2.5 ppm died Acropora sp and bleached Porites sp, Fungia sp and Pocillopora sp was avidenced within 15 days. Single exposure to cyanide at a concentration lower than that used by cyanide fisherman kills or severely impairs corals (Rubec et al., 2001), zooxanthellae density decrease upon exposure cyanide, leading to bleaching or immediate death of the zooxanthellae, but the rate of cell division increases (Cervino et al., 2003). Pocillopora damicornis colonies exposed to 4000 mg/l cyanide for 10 min causes the bleaching 90% of the coral samples within 4 days (Johannes and Riepen, 1995). Nine out of 10 Pocillopora specimens died within 4 days when exposed to CN_ concentrations of 100 mg/l for 30 min, Pocillopora specimens bleached within 3–4 days and tissue loss began after 9 days. Likewise, Pocillopora damicornis and Porites lichen were exposed to a 5200 mg/l CN_ concentration caused 100% mortality (Jones and Steven, 1997; Jones and Yellowlees, 1997) with exposure

times of 10, 20, or 30 min. Tissues coating the skeletons of Pocillopora damicornis sloughed off within 24 h. Porites lichen colonies became covered with a thick mucus-like tunic. After 7 days, the tunics of all the Porites colonies lifted off, leaving bare skeletons. Shorter exposure times (1 or 5 min) and lower CN_ concentrations (52 or 520 mg/l) induced bleaching (loss of pigment) in both coral species within 7 days (Jones and Steven, 1997). At lower concentrations resulted in the loss of zooxanthellae and impaired photosynthetic capacity, which may cause corals to die over a longer period of time (Jones and Steven, 1997; Jones and Hoegh-Guldberg, 1999; Cervino et al., 2003).

Cyanide was also shown to inhibit photosynthesis and calcification rates of Acropora formosa and A. cervicornis in corals exposed to concentrations of 5 mg/l (Chalker and Taylor, 1975). Photosynthesis and calcification were not inhibited at the highest concentration tested (5 mg/l), suggesting the existence of cyanide-resistant respiration in either the zooxanthellae or the host (Barnes, 1985).

Research by Jones and Hoegh-Guldberg (1999) showed that cyanide acts as an inhibitor of the dark reactions of the Calvin cycle; specifically, as an inhibitor of ribulose-1,5-biphosphate carboxylase/oxygenase. Jones et al. (1998) have suggested that photo-inhibition from exposure to cyanide resulted in an almost complete cessation of photosynthetic electron transport from algae in tissues of Stylophora pistillata and Acropora aspera. Cyanide was applied to small branch tips of these species at concentrations estimated to occur during cyanide fishing. Pulse-amplitude-modulation (PAM) chlorophyll fluorescence was used to examine photo-inhibition and photosynthetic electron transport in the symbiotic algae (zooxanthellae in the tissues of the corals). Therefore, cyanide fishing is regarded as one of the greatest direct threats to the sustainability of coral reefs (McManus et al., 1997; Wilkinson, 2004). As an alternative to cyanide, Erdmann (1999) suggested the use of clove oil for fishing.

207

Rahim et al./JICoR vol. 1(3) (2012): 205-216

Clove oil contains eugenol active substances derived from stems, flowers, and leaves of the clove tree (Eugenia aromatic). It has been used by humans for centuries in Indonesia as a topical anesthetic for toothaches, headaches, and joint pain (Soto and Burhanuddin, 1995). Clove oil is a naturally occurring food flavour and is extensively used in fragrance and flavor formulations for its spicy aroma. Clove oil and it’s primary ingredient eugenol have been in widespread use as flavoring and fragrance agents in the United States since before 1900. The US Food and Drug Administration (FDA) has approved clove oil or use in food as a flavoring agent, in dentistry as an analgesic and in dental cements, as a fragrance in personal care products and in aromatherapy oils, and in transdermal drug delivery systems (USPTO, 1989). Clove oil has also been found to possess antibacterial, antifungal, antiviral, antitumor, antioxidant and insecticidal properties (Chaieb et al., 2007). The FDA categorizes clove oil as generally recognized as safe (GRAS) for use in dental cement or as a food additive (US FDA, 2007).

Clove oil is also an extremely efficacious fish anesthetic, which is known to cause rapid and calm immobilization (Munday and Wilson, 1997; Keene et al., 1998.). In addition, it requires no withdrawal period after exposure (Anderson et al., 1997) and after having anesthezed, the fish will still slimy and will not loose of their appetite.

In order to use clove oil as an alternative tool for ‘eco-friendly’ fishing of ornamental fish, it should be proved that clove oil has no a harmful effect on non-target organism such as other fish and corals. There has been previous research that limited amounts of clove oil solution are unlikely to harm to coral Pocillopora damicornis (Frisch et al., 2007). There is still need to be studied on other types of coral, especially on the spesies of coral reefs in Indonesia which is dominated by Porites sp (Supriharyono, 2000). In addition, study on sub lethal effect of clove oil on the coral zooxanthellae is necessary.

Previous research has shown that corals exposed to toxic substances exhibit a variety

of responses. In the most severe cases, colonies may experience total or partial mortality, depending on the proportion of died polyps (Loya and Rinkevich, 1980; Pastorok and Bilyard, 1985). Sub-lethal effects typically include bleaching (loss of zooxanthellae and/or their pigments) or reduced photosynthetic efficiency, both of which may reduce growth, reproductive output and long-term survival (Szmant and Gassman, 1990; Jones and Hoegh-Guldberg, 1999; Cervino et al., 2003; Siebeck et al., 2006).

In its application as an alternative tool for eco-friendly fishing of ornamental fish on coral reef ecosystems, many aspects need to be examined, including the physiological response of coral animals such as changes of zooxanthellae density that can result in bleaching of coral and even can cause death of coral, hence this research needs to be done. This study aimed to analyze the effects of clove oil on changes of coral color, zooxanthellaee density and mitotic index in Porites lobata.

METHODS

Animal Test Preparation

Ninety coral colony Porites lobata sized of ± 12–15 cm in colony diameter were collected from the fringing reefs at Barranglompo Island, Makassar City, South Sulawesi on 3rd June 2011. Experimental corals were acclimated in glass tanks a week prior to the experiment. The water in the acclimatization tank was aerated and circulated.

Aquarium and Clove Oil Solution Preparation

Six experimental glass aquaria sized of 1.0x0.5x1.0 m filled with seawater with the temperature, pH and salinity of 29ºC, 7.8 and 32 mg/L, respectively and completed with aeration were used in this experiment. Stock solution of clove oil was prepared by diluting clove oil in ethanol with ratio of 1:8. Eugenol content in the clove oil was analyzed by HPLC method that was 83.058%.

208


This stock solution was diluted evenly into the five experimental aquarium based on the concentration treatments. There were five treatment concentrations of clove oil, i.e. 20, 30, 40, 50 and 60 ppm and one as a control with no addition of clove oil.

Experiment Design

This experiment was conducted to observe the effects of clove oil on the color changes, zooxanthellae density and mitotic index of the experimental coral. Experimental design was a factorial design, which consisted of six concentration (control, 20, 30, 40, 50, 60 ppm) and five time (1, 5, 9, 13, and 17 days) treatments with 3 replication. Steps of this experiment as follows:1). Coral was placed into the aquarium that

contains clove oil solution of different concentrations (control, 20, 30, 40, 50, 60 ppm) for 5 min and then transferred into a tank containing clean seawater for 60 seconds and then removed to other tanks for recovery process. Water in the recovery tank were aerated and circulated.

2). For recovery process, observation was made everyday for coral color changes for 17 days period and the color changes was score based on the standard “coral health chart” (Coral Watch, the University of Queensland) and for zooxanthellae density and mitotic index observation on zooxanthellae density and mitotic index were carried-out at 1, 5, 7 and 13 days after exposure to clove oil.

Zooxanthellae density in coral was calculated based on Suharsono and Sukarno (1983). Zooxanthellae was taken by spraying the coral surface 1 cm2 with compressed air, filtered with 25 micrometers sieve then it was diluted into 100 mL sea water. Number of zooxanthellae was counted by haemocytometer and its density was calculated using the formula APHA (1992) which has been modified by Niartiningsih (2001), as follows:

where: N = number of organisms counted (cell/cm2); At = area of cover glass (mm2); Vt = volume of total initial sample (ml); Ac = area of sprayed samples (cm2); Vs = volume of sample used (ml); As = area of micrometer (mm2).

Observation on the mitotic index of zooxanthellae followed Suharsono (1990), Brown and Zamani (1992), Zamani (1995), and Ambariyanto (1996) by homogenization method that zooxanthellae was taken from the coral by spraying strong air generated by mini compressor on the coral and the fallen soft tissue was collected in the transparent plastic bag. The sample, then was suspended in the clean seawater. To separate between zooxanthellae and debris, the sample was filtered in series of filters sized of 250, 175 and 50 mm. The filtered sample was placed in volumetric glass and was ready to be analyzed. Mitotic index was determined by counting zooxanthellae in the phase of cytokinesis or karyokinesis using a Neubauer haemocytometer which was appears as twin cells on the microscope with 400 magnification. This observation was conducted during 24 hrs at 03.00, 06.00, 09.00, 12.00, 15.00, 18.00, 21.00 and 24.00 hrs with five replication. Mitotic index was determined based on Ambariyanto (1996) with equation of:

Data Analysis

Analysis of data obtained from the experiments was performed using Microsoft Office Excel and SPSS software. Normality of data was tested with the Kolmogorov-Smirnov Test and Shapiro-Wilk test, while the homogeneity of variance was tested by Levene’s Test. Test the significance of the different levels of data carried by two factorial ANOVA test and Post HocTest with Tukey HSD Test. The data are presented in tables and graphs.

209


RESULTS AND DISCUSSION

Change of Coral Color

Observation on the coral colour change after exposure to clove oil are presented in Table 1. It can be seen from the table that the score value of coral exposed to treated concentration of clove oil is 5.5. This indicates the coral is in healthy condition.

Similar results was also found by Frisch et al. (2007) that coral Pocillopora damicornis exposed to 0.5 ppt of clove oil for 60 minutes produce health score value and it did not differ significantly from the control corals. He found also that exposing the coral at higher concentrations of clove oil (5 ppt) for 1 minute resulted healthy coral and the score value is not significantly different from the control corals.

Table 1. Coral Health Value in Porites lobata that observed for 17 days after exposure to some concentration of clove oil.

Time of Exposure (Day-)

Concentration of Clove Oil (ppm)

Control 20 30 40 50 60

Before 5.5 5.5 5.5 5.5 5.5 5.5

After 5.5 5.5 5.5 5.5 5.5 5.5

1 5.5 5.5 5.5 5.5 5.5 5.5

2 5.5 5.5 5.5 5.5 5.5 5.5

3 5.5 5.5 5.5 5.5 5.5 5.5

4 5.5 5.5 5.5 5.5 5.5 5.5

5 5.5 5.5 5.5 5.5 5.5 5.5

6 5.5 5.5 5.5 5.5 5.5 5.5

7 5.5 5.5 5.5 5.5 5.5 5.5

8 5.5 5.5 5.5 5.5 5.5 5.5

9 5.5 5.5 5.5 5.5 5.5 5.5

10 5.5 5.5 5.5 5.5 5.5 5.5

11 5.5 5.5 5.5 5.5 5.5 5.5

12 5.5 5.5 5.5 5.5 5.5 5.5

13 5.5 5.5 5.5 5.5 5.5 5.5

14 5.5 5.5 5.5 5.5 5.5 5.5

15 5.5 5.5 5.5 5.5 5.5 5.5

16 5.5 5.5 5.5 5.5 5.5 5.5

17 5.5 5.5 5.5 5.5 5.5 5.5

210


In this study also looks Porites lobata coral color at all exposure concentrations (20–60 ppm) observed there were no color changes when compared with control corals. Even up to the 17th day after exposure to the highest concentration (60 ppm) was not find any color changes of the coral (Fig. 1). This indicates that clove oil does not have a significant influence on coral color changes a concentration of 20–60 ppm. This means that the use of clove oil at a concentration of 20–60 ppm has not led to changes in the health of the coral. These results are consistent with a previous study by Frisch et al. (2007) which states that exposure to clove oil at a concentration of 0.5 ppt does not affect the color change of Pocillopora damicornis. Further said that clove oil exposure with a concentration of 5 ppt not influence the photosynthetic capacity of the coral Pocillopora damicornis.

Things are different on cyanide exposure was done by Subandi (2004), where the corals exposed to cyanide concentration of 2.5 ppm has undergone changes in color (bleaching) starting on day 1 after exposure to all types of coral (Acropora, Pocillopora, Porites and Fungia), but on day 7 to day 13 after exposure, it appears corals have undergone a process of recovery, unless the Acropora coral, recovery process does not occur at all that coral bleaching remained until the end of the study (day 15).

The same study has also been undertaken by the Jones and Steven (1997) which reported the effect of exposure to cyanide at various levels of concentration of coral species Pocillopora damicornis. Exposure to concentrations of 0.4% for 10 minutes causes the bleaching 90% of the coral samples within 4 days. Exposure to concentrations of 0.01%, bleaching begin within 3–4 days and the loss of coral tissue showed after 9 days (Johannes and Riepen, 1995). Jones and Steven (1997) also reported that exposure to cyanide to zooxanthellae coral symbiosis in the level as occurs in cyanide fishing caused bleaching or the death of coral polyps.

Research Cervino et al. (2003) on the coral Acropora, Millepora, Heliofungia sp and Trachyphyllia geoffrio were exposed to cyanide produce color changes in corals, where bleached Acropora millepora within 24 hours, Heliofungia bleached within 72 hours after exposure and bleached, whereas Trachyphyllia geoffrio 3 weeks after exposure.

Density of Coral Zooxanthellae

In addition to observe the color changes in corals, the study also analyzed the coral zooxanthellae density. It aims to look as far as possible the effect of clove oil on the coral, including the zooxanthellae. According to Nganro (2009), the phenomenon that

Fig 1. Coral color of Porites lobata sp: (a) Before exposure to clove oil (control). (b) Day 17 after exposure to 60 ppm of clove oil.

211


occurs due to the loss zooxanthellae release off Coelenterate because of the stress. Zooxanthellae loss is generally regarded as a response to stress conditions. It can be induced by storms, changes in lighting, low salinity, temperature changes and pollution.

Quick release of zooxanthellae can occur directly from endodermal cells towards coelenteron (Suharsono, 1990), there may be certain biochemical response that induces cell contraction in host cells which may zooxanthellae lost quickly, especially when the host is in stress (Nganro, 2009).

Based on the analysis conducted after exposure to several concentrations of clove oil (20, 30, 40, 50, 60 ppm) in Porites lobata, the results obtained can be seen in Table 2 and Fig. 2. Anova test results on the error level of 5% (α = 0.05) indicated that there was no significant difference of zooxanthellae density in Porites lobata on the clove oil concentration sprayed (P> 0.05), while the variation of time (days) were seen significantly difference (P <0.05). In addition, the test also showed that there is no interaction between the two treatment variables (concentration and time) (P> 0.05).

Post Hoc test of time variation with Tukey HSD was done to determine which time is different. The results showed that the zooxanthellae density of the Porites lobata

on day-1 was different to day 5, 9, 13 and 17, where the zooxanthellae density on day-1 was less than that on the next day (5th, 9th, 13th and 17th). This shows that there has been an increase in the density zooxanthellae of the Porites lobata after 1 day recovery period from clove oil exposure. This is very different from the coral under pressure due to changes in the environment which will decrease the zooxanthellae density, as has been reported by Cervino et al. (2003) which states that the number of coral zooxanthellae decreased after exposure to cyanide for 120 seconds. Research conducted by Nganro (2009) on Anemonia viridis exposured some concentrations copper also showed zooxanthellae release of its host during the 5 days, where the control condition remained stable.

The density of zooxanthellae of Porites lobata after clove oil exposured at several concentrations was seen no significant change (Table 2). This can be proved by observing the zooxanthellae density on coral treatments (exposed clove oil) with a control coral (without clove oil treatment) saw no difference. Zooxanthellae density observed ranged from 3.43 to 8.47 x 106 cells/cm2 at all concentrations of clove oil were observed, as well as control corals. This is also evidenced by the statistical test (P> 0.05) in which there was no significant difference between the density of zooxanthellae on

Table 2. The average of zooxanthellae density in Porites lobata after exposure to some concentrations of clove oil.

Concentrations of Clove Oil (ppm)

Changes of Zooxanthellae Density ( x 106 cell/cm2)

Day 1 Day 5 Day 9 Day 13 Day 17

Control 3.43 5.97 5.60 6.93 5.77

20 4.80 7.40 7.20 5.70 6.23

30 3.67 5.53 5.77 5.70 7.00

40 4.20 5.33 5.93 5.37 7.57

50 4.00 5.80 6.50 6.83 8.47

60 4.00 5.67 5.67 5.40 5.50

212


Porites lobata with several concentrations of clove oil is sprayed. However, observation of zooxanthellae density at various time series statistically showed significant difference. The zooxanthellae density fluctuated after a few days of observation, whether on the coral controls or corals exposed to clove oil. Density was highest at day 13 and 17 after exposure in concentrations of 40 ppm and 50 ppm respectively.

Zooxanthellae density in the coral endo-dermic tissue was varied, depending on the type of host and environmental factors. In a normal water condition, zooxanthellae in the coral tissue fluctuates within normal limits. This is an indicator of balance con-dition between growth of coral tissue and zooxanthellae (Suharsono, 1992).

Density of coral zooxanthellae was various. Drew (1972), Muscatine et al. (1985), and Harland and Brown (1989) was 0.5–8.5x106 cells/cm2, depending on species and habitat depth, where the zooxanthellae host is taken, whereas Niartiningsih (2001) indicated the average zooxanthellae density of the coral Acropora samoensis was 2.74x106 cells/cm2. Observation on day 5, 9, 13 and 17 after exposure of clove oil showed an increase trend in the zooxanthellae density even though small fluctuation occurred from day 1 to 17 (Fig. 2). In the contrary, 120 second

cyanide exposure to various corals, their zooxanthellae density decreased since 1 to 30 days after exposure (Cervino et al., 2003). This indicates clove oil is safe to the corals within these treated concentrations.

Mitotic Index of Coral Zooxanthellae

Mitotic index is the cell division rate which is expressed as a percentage of cells undergoing division in the zooxanthellae population. Rate of cell division or mitotic in symbiotic zooxanthellae with invertebrates are very sensitive to environmental changes (Gladfelter, 1985).

In the symbiotic algae in some Coelenterata, mitotic index of zooxanthellae has been used to measure changes of algae response to natural and experimental conditions (McAuley, 1985). Mitotic index and growth rate of symbiotic zooxanthellae from various host known to be influenced by the concentration of nutrients (Wilkerson et al, 1988). Another factor affecting the mitotic index is irradiation (McAuley, 1985), depth (Wilkerson et al., 1988), temperature (Gates and Brown, 1985) and the treatment of crude oil to the host (Michel and Fitt, 1984).

Observations on the mitotic index coral Porites lobata can be seen in Table 3. Test of normality (Kolmogorof-Smirnov and Shapiro-Wilk) and test of variance (Levene’s

Fig 2. Zooxanthellae density in Porites lobata sp after exposure to several concentration of clove oil.

213


test) on the zooxanthellae mitotic index of coral Porites lobata against variations in the concentration and time (days) indicated that the data does not meet the requirements of normality and homogeneity of variance although have been transformed so the data was statistically analyzed by non-parametric test (Kruskal-Wallis test). The results showed that zooxanthellae mitotic index on the coral exposed to the all treatment with various concentrations of clove oil exposure was not significantly different, whereas mitotic index based on the variation of time (days) showed significantly different.

Observations of zooxanthellae mitotic index on Porites lobata exposed to clove oil has showed no different from the control. This suggests that exposure to clove oil to a concentration of 20 to 60 ppm does not cause changes in coral zooxanthellae mitotic index. However, in all treatment and control corals mitotic index fluctuated during the exposure period from day 1 to 17 but this fluctuation was still in acceptable level. This indicates that cell division continued during the exposure period as seen in Fig. 3.

Different result was reported by Cervino

Table 3. The average of zooxanthellae mitotic index in Porites lobata after exposure to some concentrations of clove oil.

Concentration of Clove Oil (ppm)

Change of Zooxanthella Mitotic Index (%)

Day 1 Day 5 Day 9 Day 13 Day 17

Control 0.280 0.270 0.270 0.290 0.280

20 0.242 0.250 0.280 0.293 0.283

30 0.270 0.285 0.297 0.278 0.262

40 0.260 0.285 0.255 0.275 0.277

50 0.243 0.262 0.277 0.277 0.282

60 0.238 0.265 0.257 0.285 0.268

Fig 3. Zooxanthellae mitotic index in Porites lobata after exposure to several concentration of clove oil.

214


et al. (2003) that mitotic index increased after exposure the coral Euphyllia divisa to cyanide at a concentration of 600 mg/L and 50 mg/L. Zooxanthellae in controls divided approximately two times less frequently than those in corals exposed to 50–600 mg/L cyanide. Rapid increasing in mitosis leading to expulsion of zooxanthellae were also evident during their experiment. This can leave the coral with reduced photosynthetic capabilities. Continuous cell division of algal populations may result from lower densities of zooxanthellae, condition that may induce density-dependant division in order to restore original algal concentrations. A similar outcome occured in experiment Jones and Yellowlees (1997) when coral were exposed to various concentrations of copper. Nganro (2009) reported exposure the Anemonia viridis to copper at a concentration of 0,05 mg/L also showed increasing mitotic index of zooxanthellae after 30 min treatment.

CONCLUSIONThe results of this study indicate that the

use of clove oil at concentration of 20–60 ppm has no negative effect on the coral in the aspects of coral color change, density and mitotic index of corals zooxanthellae. Therefore, clove oil is considered safe as an alternative tool for eco-friendly fishing of ornamental fish in coral reef.

REFERENCES

Ambariyanto. 1996. Effect of Nutrient Enrichment in the Field on the Giant Clam, Tridacna maxima. PhD Thesis. School of Biological Science and Marine Studies Centre, the University of Sydney, Australia. 269 p.

Anderson, W.G., McKinley, R.S., and Colavecchia, M. 1997. The use of clove oil as an anesthetic for rainbow trout and its effects on swimming performance. North American Journal of Fisheries Management, 17(2): 301–307.

Barnes, D.J. 1985. The effects of photosynthetic and respiratory inhibitors upon calcification in the staghorn coral Acropora formosa. Proceedings of the 5th International Coral Reef Congress, 6: 161–166.

Brown, B.E. and Zamani, N.P. 1992. Mitotic indices of zooxanthellae: A comparison of technique based on nuclear and cell division frequencies. Mar. Ecol., 89: 99–102.

Cervino, J.M., Hayes, R.L., Honovich, M., Goreau, T.J., Jones, S., and Rubec, P.J. 2003. Changes in zooxanthellae density, morphology, and mitotic index in hermatypic corals and anemones exposed to cyanide. Mar. Pollut. Bull., 46: 573–586.

Chaieb, K., Hajlaoui, H., and Zmantar, T. 2007. The chemical composition and biological activity of clove essential oil, Eugenia caryophyllata (Syzigium aromaticum L. Myrtaceae): A short review. Phytotherapy Res, 21: 501–506.

Chalker, B.E. and Taylor, D.L. 1975. Light-enhanced calcification, and the role of oxidative phosphorylation in calcification of the coral Acropora cervicornis. Proceedings of the Royal Society of London Series B, 190: 323–331.

Drew, E.A. 1972. The biology and physiology of alga–invertebrate symbioses. II. The density of symbiotic algal cells in a number of hermatypic hard corals and alcyonaceans from various depths. J. Exp. Mar. Biol. Ecol., 8: 71–75.

Erdmann, M.V. 1999. Clove oil: an ‘eco-friendly’ alternative to cyanide use in the live reef fish industry? SPC Live Reef Fish Bulltetin, 5: 4–7.

Frisch, A.J., Ulstrup, K.E., and Hobbs, J.A. 2007. The effects of clove oil on coral: an experimental evaluation using Pocillopora damicornis. Journal of Experimental Marine Biology and Ecology, 345: 101–109.

Gates, R. and Brown, B.E. 1985. The ‘loss’ of zooxanthellae in the sea anemone Anemonia viridis (Forsskal) under stress. Proc. 5th Int. Coral Reef Congr. 2: 143.

Gladfelter, E.H. 1985. Metabolism, calcification and carbon production-organism level studies. Proc. 5th Int. Coral Reef Congr.Tahiti, 4: 527–537.

Harland, A.D. and Brown, B.E. 1989. Metal tolerance in the scleractinian coral Porites lutea. Mar. Pollut. Bull., 20: 353–357.

Johannes, R.E. and Riepen, M. 1995. Environmental, Economic, and Social Implications of the Live Reef Fish Trade in Asia and the Western Pacific. The Nature Conservancy, Honolulu.

215


Jones, R.J. and Steven, A.L. 1997. Effects of cyanide on corals in relation to cyanide fishing on reefs. Mar. Freshw. Res., 48: 517–522.

Jones, R.J. and Yellowlees, D. 1997. Regulation and control of intracellular algae (zooxanthellae) in hard corals. Phil. Trans. Royal Society London, 352: 457–468.

Jones, R.J., Hoegh-Guldberg, O., Larkum, A.W.D., and Schreiber, U. 1998. Temperature-induced bleaching of corals begins with impairment of the CO2 fixation mechanism in zooxanthellae. Plant Cell Environ., 21: 1219–1230.

Jones, R.J. and Hoegh-Guldberg, O. 1999. Effects of cyanide on coral photosynthesis: implications for identifying the cause of coral bleaching and for assessing the environmental effects of cyanide fishing. Marine Ecology Progress Series, 177: 83–91.

Keene, J.L., Noakes, D.G., Moccia, R.D., and Soto, C.G. 1998. The efficacy of clove oil as an anaesthetic for rainbow trout, Oncorhynchus mykiss (Walbaum). Aquaculture Research, 29: 89–101.

Loya, Y. and Rinkevich, B. 1980. Effects of oil pollution on coral reef communities. Mar. Ecol. Prog. Ser., 3: 167–180.

McAuley, P.J. 1985. The cell cycle of symbiotic Chlorella I. The relationship between host finding and algal cell growth and division. Journal Cell Sci., 77: 225–239.

McManus, J.W., Nañola, C.L., and Reyes, R.B. 1997. Effects of some destructive fishing methods on coral cover and potential rates of recovery. Environmental Management, 21(1): 69–78.

Michel, W.C. and Fitt, W.K. 1984. Effects of a water-soluble fraction of a crude oil on the coral reef hydroid Myrionema hargitii: feeding, growth and algal symbiont. Mar. Biol., 84: 143–154.

Munday, P.L. and Wilson, S.K. 1997. Comparative efficacy of clove oil and other chemicals in anaesthetization of Pomacentrus amboinensis, a coral reef fish. Journal Fish Biology, 51: 931–938.

Muscatine, L., McCloskey, L.R., and Loya, Y. 1985. A comparison of the growth rates of zooxanthellae and animal tissue in the Red Sea coral Stylophora pistillata. Proc. 5th Int. Coral Reef Symp., 6: 119–123.

Nganro, N.R. 2009. Metoda Ekotoksikologi Perairan Laut Terumbu Karang. Sekolah

Ilmu dan Teknologi Hayati, Institut Teknologi Bandung, Bandung.

Niartiningsih, A. 2001. Analisis Mutu Zooxanthellae dari Berbagai Inang dan Pengaruhnya terhadap Sintasan dan Pertumbuhan Juvenil Kima Sisik (Tridacna squamosa). Disertasi. Program Pascasarjana Universitas Hasanuddin, Makassar.

Pastorok, R.A. and Bilyard, G.R. 1985. Effects of sewage pollution on coral-reef communities. Mar. Ecol. Prog. Ser., 21: 175–189.

Rubec, P.J., Cruz, F., Pratt, V., Oellers, R., McCullough, B., and Lallo, F. 2001. Cyanide-free net-caught fish for the marine aquarium trade. Aquar. Sci. Conserv., 3: 37–51.

Siebeck, U., Marshall, N.J., Kluter, A., and Hoegh-Guldberg, O. 2006. Monitoring coral bleaching using a colour reference card. Coral Reefs, 25: 453–460.

Soto, C.G. and Burhanuddin. 1995. Clove oil as a fish anaesthetic for measuring length and weight of rabbitfish (Siganus lineatus). Aquaculture, 135: 149–152.

Subandi, N. 2004. Pengembangan Metode Penyidikan Ilmiah Untuk Pembuktian Kasus-kasus Penangkapan Ikan dengan Bahan Peledak dan Sianida. Disertasi. Sekolah Pascasarjana IPB, Bogor.

Suharsono. 1990. Ecological and Physiological Implication of Coral Bleaching at Pari Island, Thousand Island, Indonesia. Dissertation. University of New Castle Upon Tyne, UK.

Suharsono. 1992. Ultra structure of the endosymbiotic dinoflagellate Symbiodinium microadriaticum living in sea anemon. Mar. Res. in Indonesia, 28: 13–33.

Suharsono and Sukarno. 1983. Kandungan zooxanthellae pada karang batu di terumbu karang Pulau Pari. Pewarta Oseana, 16: 1–7.

Supriharyono. 2000. Pengelolaan Ekosistem Terumbu Karang. Penerbit Djambatan, Jakarta.

Szmant, A.M. and Gassman, N.J. 1990. The effects of prolonged bleaching on the tissue biomass and reproduction of the reef coral Montastrea annularis. Coral Reefs, 8: 217–224.

US FDA. 2007. Guidance for Industry: Concerns Related to the Use of Clove Oil as an Anesthetic for Fish. Guideline No. 150. US Food and Drug Administration Center for Veterinary Medicine. http://www.fda.gov/cvm/Guidance/guide150. htm.

USPTO. 1989. Eugenol enhancement of transdermal drug delivery. US Patent 4888362.

216


Wilkerson, E.P., Kobayashi, D. and Muscatine, L. 1988. Mitotic index and size of symbiotic algae in Caribbean reef corals. Coral reefs, 7(1): 29–36.

Wilkinson, C. 2004. Status of Coral Reefs of the World: Volume 2. Australian Institute of Marine Science, Townsville, Queensland, Australia. 301 p.

Zamani, N.P. 1995. Effect of Environmental Stress on Cell Division and Other Celluler Parameters of Zooxanthellae in the Tropical Symbiotic Anemone Heteractis malu, Haddon and Shackleton. Dissertation. Marine Science and Coastal Management Department, University of New Castle Upon Tyne, United Kingdom. 261 p.

The Effects of Clove Oil in Color Changes and ... · The Effects of Clove Oil in Color Changes and...

Documents

Transcript of The Effects of Clove Oil in Color Changes and ... · The Effects of Clove Oil in Color Changes and...