The anti-leech potential of Turmeric ( Curcuma domestica ) Juice against Sea Leech in Cantang Hybrid Grouper ( Epinephelus fuscoguttatus >< Epinephelus lanceolatus )

. One of the diseases that attacks and disrupts productivity in hybrid grouper cultivation (Epinephelus fuscoguttatus >< Epinephelus lanceolatus) is caused by the sea leech parasite ( Zeylanicobdella sp.). To control parasite attacks, safe and easy control measures need to be taken, one of which is the application of turmeric juice to soak fish infected with leeches. Therefore, research must be conducted to test the most effective concentration and soaking time of turmeric ( Curcuma domestica ) juice to reduce the intensity of sea leech attacks on hybrid grouper fish. This study was to test differences in turmeric juice concentration using a completely randomized factorial design consisting of 2 factors, namely the turmeric juice concentration factor (0 mL/L, 0.6 mL/L, 0.9 mL/L and 1.2 mL/L seawater) and immersion time factor (30 and 60 minutes) with eight treatments and three replications. The research results showed that the most effective reduction in intensity after soaking with turmeric juice was at a concentration of 1.2 mL/L of seawater for 60 minutes at 24.00%. The interaction between the concentration and soaking time of turmeric juice reduced the intensity of parasites but did not affect the behaviour and survival of the fish. Turmeric juice has the potential as an anti-leech and can be applied in grouper cultivation to reduce the intensity of leech infections.


Introduction
Cantang grouper is the result of a cross between male giant grouper (Epinephelus lanceolatus) and female tiger grouper (Epinephelus fuscoguttatus), which was produced to develop a new grouper variety that has several benefits in terms of nutrition, growth and economic value.This fish has twice the growth advantage compared to other grouper fish generally cultivated by the community [1].The "Cantang" hybrid grouper (Epinephelus fuscoguttatus >< Epinephelus lanceolatus) is a superior aquaculture commodity that has local and international market potential.One of the challenges in grouper cultivation is the emergence of disease attacks, which cause a decrease in aquaculture production.Fish death is usually caused by pathogenic infections such as parasites, fungi, bacteria and viruses, which can cause disease [2].One of the diseases that attack fish is caused by parasites, which are detrimental to the fish, thereby disrupting the productivity of the cantang grouper.Parasitic diseases can reduce the productivity and quality of the fish, thereby causing financial losses for producers.Leeches are a common parasite that attacks cantang grouper fish [3].Sea leech infection in farmed fish can hamper the development and health of fish because this parasite sucks the blood of its host (fish).The sea leech parasite (Zeylanicobdella sp.) is a parasite that infects various types of grouper fish, including cantang hybrid grouper [4].
Treatment of ectoparasite infections can be done chemically and using natural ingredients.Chemical treatment can have a harmful impact because it is carcinogenic and expensive to fish.Apart from that, using natural materials has advantages such as being relatively safer, not causing resistance, cheap and not harmful to the surrounding environment [5].Parasitic diseases can be prevented by using herbal components; one of the herbs that can be used is turmeric.Turmeric is known as a plant that has antiparasitic properties.Turmeric contains active metabolites such as turmerone, zingiberin, felandrene, phenolics, and curcumin, which help cure parasitic infections.Besides being antiparasitic, turmeric has anti-inflammatory effects due to its polyphenol content [6].Turmeric has been proven to reduce and inhibit the growth of ectoparasites in the hatching media for gourami fish (Osphronemus gouramy) eggs [7].The anti-leech potential of turmeric juice has yet to be discovered.Therefore, it is necessary to research turmeric juice to reduce the intensity of leeches in cantang grouper fish.The role of turmeric can be applied through soaking.This research aims to test the concentration of turmeric juice (Curcuma domestica) that is most effective in reducing the number of sea leech ectoparasites in cantang grouper fish using the soaking method.

Preparation of Turmeric Juice
The turmeric sample used as test material is from the Bintan Center market, Tanjungpinang.Making turmeric juice [9], which begins with washing the turmeric rhizomes until clean; the turmeric is chopped into small pieces and ground, then filtered using filter paper to extract pure juice.

Preparation of Containers and Media
The containers needed are 24 round buckets with a capacity of 16 litres.Fill 8 liters of seawater into the rearing container, the sea water is aerated before soaking the "cantang" hybrid grouper with turmeric juice.

Immersion of "Cantang" Hybrid Grouper Treatment Experiment
This research was carried out with a completely randomized factorial design, which consisted of the factors of turmeric juice concentration (0 mL/L, 0.6 mL/L, 0.9 mL/L and 1.2 mL/L seawater) and immersion time (30 and 60 minutes) on fish infected with marine leeches.Immersion "cantang" hybrid grouper fish is done by adding turmeric juice according to a predetermined concentration to a soaking container that has been previously aerated.After mixing the turmeric juice, put 1 test fish into the container.The intensity of leeches was observed before immersion using a squeeze with direct observation to see the number of fish attacked by parasites and the number of parasites that attacked.

Immersion of "Cantang" Hybrid Grouper Treatment Experiment
This research was carried out with a completely randomized factorial design, which consisted of the factors of turmeric juice concentration (0 mL/L, 0.6 mL/L, 0.9 mL/L and 1.2 mL/L seawater) and immersion time (30 and 60 minutes) on fish infected with marine leeches.Immersion of "cantang" hybrid grouper fish is done by adding turmeric juice according to a predetermined concentration to a immersion container that has been previously aerated.After mixing the turmeric juice, put 1 test fish into the container.The intensity of leeches was observed before immersion using a squeeze with direct observation to see the number of fish attacked by parasites and the number of parasites that attacked.Fish survival was observed before and after immersion treatment.The swimming behaviour of the fish was observed during immersion.

Data Analysis
Data on parasite intensity and fish survival were analyzed using Two-Way ANOVA (Analysis of Variance).If the treatment had a significant impact, further tests were carried out with SPSS IBM 25 to determine the natural effect of therapy on sea leeches (Zeylanicobdella sp.).Fish behaviour was analyzed descriptively.

Sea Leech Intensity
The average percentage reduction in intensity (%) after immersion treatment is presented in Table 1.The interaction between the concentration given and the immersion time used influenced the percentage reduction in intensity of Zeylanicobdella sp.The best concentration of turmeric juice to reduce the intensity percentage of Zeylanicobdella sp. in "cantang" hybrid grouper was 1.2 mL/L seawater, and this concentration treatment was significantly different from the concentration treatment of 0.9 and 0.6 mL/L seawater and the control (treatment without turmeric juice).Apart from that, the best immersion time of turmeric juice is to reduce the intensity of Zeylanicobdella sp. in "cantang" hybrid grouper, namely in the treatment with an immersion time of 60 minutes, which treatment was significantly different (P<0.05) to the immersion time of 30 minutes.Note : Different letter in the supescript showed the significant different between the treatment (P<0.05).

Survival Rate of "Cantang" Hybrid Grouper
The results of survival observations in "cantang" hybrid grouper after the immersion treatment can be seen in Table 2.The survival rate of "cantang" hybrid grouper in all treatments was 100 ± 0.00%.The immersion treatments did not significantly differ (P>0.05) in the survival of the "cantang" hybrid grouper.Note : (+) : significant different between concentration and duration of immersion (P>0.05).

"Cantang" hybrid Grouper Swimming Behavior
"Cantang " hybrid grouper swimming response to immersion treatments given showed no difference in changes in behavior at each concentration and immersion time from pretreatment, treatment and post-treatment.Fish swam actively in all treatments.

Discussion
Intensity is the average number of parasites found from the number of infected fish.Intensity shows how much the ectoparasite attacks [10].The results of intensity observations in each treatment showed that the average intensity of sea leeches decreased after soaking in a solution of turmeric juice.The best reduction in intensity was found at a concentration of 1.2 mL/L with a soaking time of 60 minutes, amounting to 24.00%.The results of the research show that the higher the soaking concentration and the longer the immersion time, the more optimal the ability of turmeric juice to reduce sea leech infections in "cantang" hybrid grouper.Turmeric has anti-parasitic properties.Turmeric is effective in reducing the intensity of ectoparasites in tilapia because it contains ingredients such as curcumin, flavonoids, essential oils and tannins, which can disrupt the development of the nervous system, damage the nervous and respiratory systems, and reduce the ability of parasites to digest food [11].Reducing the intensity using turmeric juice was more effective than using turmeric extract, with a reduction in the leech intensity percentage of 21.88% [11].Turmeric juice is also more effective than the use of other herbal ingredients, such as garlic powder with a reduction in the percentage intensity of leeches, namely 18.45% [12], noni leaf extract with a decrease in the percentage intensity of ectoparasites of 0.30% [13].However, turmeric juice is less effective than the chemical cupric sulfate (CuSO4), with a 100% reduction in sea leech intensity [14].
Parasites in fish generally cause harmful effects [15].Parasite attacks make the fish lose their appetite, then slowly become weak and lead to death.Clinical symptoms caused by infestation of sea leeches Zeylanicobdella sp. are that the fish experiences a decrease in appetite, swims weakly on the surface of the water, and the fish experiences swelling of the skin.Heavily-intensity fish show symptoms of being quiet and weak at the bottom of the rearing tank [5].Fish infested with sea leeches Zeylanicobdella sp. have visible bleeding, swelling around the anterior and posterior sucking areas on the host's skin and lesions characterized by white ulcers on the surface of the host's skin [16].
Fish survival can be defined as the chance of living at a specific time [17].Based on research, the survival rate of "cantang" hybrid grouper infected with parasites has a high chance of survival because the S.R. value reaches 100%.The treatment concentration used during the research was still relatively safe.Therefore, when soaking turmeric juice with a concentration of up to 1.2 mL/L of seawater, the fish can still adapt to the attention given and do not cause abnormal changes in their swimming response.
The results of observations during research on "cantang" hybrid grouper infested with Zeylanicobdella sp.show normal swimming behaviour both before, during, and after immersion.Apart from that, after soaking the turmeric it was given, it showed that the condition of the fish's swollen body began to decrease.When turmeric juice was given, the fish's behaviour did not change.It continued to move generally because the fish adapted to the concentration and length of soaking turmeric juice given.Turmeric juice can be applied by soaking fish infected by leeches because it is safe and can reduce the intensity of leeches by 21.88%.

Conclusion
Soaking in turmeric juice (Curcuma domestica), which was effective in this study in reducing the intensity of sea leeches (Zeylanicobdella sp.) in cantang grouper (Epinephelus fuscoguttatus >< Epinephelus lanceolatus) was at a concentration of 1.2 mL/L of seawater with a soaking time of 60 minutes, which can reduce the leech intensity by 24.00%.Fish farmers can use turmeric juice to reduce the intensity of leeching through the soaking method.

Table 1 .
The average percentage reduction in intensity (%) after immersion treatmen.

Table 2 .
Survival rate of "cantang" hybrid grouper after the immersion treatment.