Effects of Honey-Induced Sex Reversal on Sex Ratio and Growth Performance of the Nile Tilapia (Oreochromis niloticus)

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1 Nigerian Journal of Fisheries and Aquaculture Nigerian Journal of Fisheries and Aquaculture 6(1):1 6 May, 2018 Copy, Right 2013 Printed in Nigeria. All rights of reproduction in any form reserved. Department of Fisheries, Faculty of Agriculture, University of Maiduguri, Nigeria Available on line: ISSN Nig. J. Fish. Aquac. Effects of Honey-Induced Sex Reversal on Sex Ratio and Growth Performance of the Nile Tilapia (Oreochromis niloticus) Abdullah, N. Department of Aquaculture, Faculty of Fisheries and Marine Science, Universitas Khairun Ternate Indonesia Received March 20, 2017 Accepted: May 27, 2017 ABSTRACT Sex reversal is one of the techniques used to improve growth in fish and it is traditionally achieved using synthetic material such as hormones. However, the use of synthetic hormones is not common among fish breeders in North Moluccas. Other synthetic material such as honey, in fact, can be used to do so (Honey is an organic substance not synthetic). Honey is a natural substance that can be used to reverse the sex of fish as it contains potassium and chrysene that act as the natural aromatase inhibitors. In this research, four (4) treatments concentration of 0.0 (control) 1.0, 1.5, and 2.0 of honey in 2L of water were tested to determine its effects on sex ratios and growth of the Nile Tilapia, Oreochromis niloticus. The 7-day old larva was immersed in the solution for 2 hours. The research design used was the Completely Randomized Design (CRD). The data obtained was analysed using the variance analysis at the confidence level at 99 % and followed by a BNT test to see the effect among treatments on the tested parameters. The result showed that administration of honey resulted in a significantly (p>0.05) different percentage of male sex in comparison to the control while there was no significant difference among honey dosage. The highest number of males (86%), however, obtained in treatment of while in treatment B with the dosage of 1.5% males was 84%. In contrast, the lowest percentage was found in the treatment D with the dosage of 0% (control) that was 38%. The survival rates in the treatment given with the dosage of honey A (1.0%), B (1,5%), and C (2,0%) were significantly different from the treatment D without any dosages of honey (0%). The growth was not significantly different from others. Key words: Honey, Sex reversal, Oreochromis niloticus, male, immersion INTRODUCTION The success in a fish culture can be seen from fish production with a rapid growth in a shorter period. The target of production can be in the form of total number of fish yielded by measuring the survival rate - particularly for the hatchery and can be in the form of weight obtained by calculating the biomass in the rearing and growth process. Synthetic hormone has been used to produce all fish through sex reversal technique by using hormone through immersion, injection and feeding. For the purpose of sex reversal, it is more preferred to use the hormone of 17α-methyl-testosterone as it theoretically can be expected to yield the population of 100% of male (Zairin, 2002). Sex reversal is one of the techniques used to improve growth in fish and it is traditionally achieved using synthetic material such as hormones. Honey is a natural substance that can be used to reverse the sex of fish as it contains potassium and chrysene that act as the natural aromatase inhibitors. Several studies authors reported 100% male tilapia after treating with 17α-methyl-testosterone (Pandian and Varadaraj, 1988; Afanso et al., 2001; Mubarik, 2011). Mansori et al. (2004) documented 96% males Thai red Tilapia fed with fluoxymesteron based diet. The use of natural hormone for sex reversal have also been documented some authors; Haylor and Pascal (1991) reported successful sex reversal using ram testes. Chukwu et al. (2013) obtained 92.86% male O. niloticus after feeding with a papaw-based diet for 30 days. Recently, immersion using a number of synthetic hormones have been conducted, faced disadvantages of being carcinogenicity and potential waste for the environmental pollution. Hence, purpose of this nrs.abdulla@gmail.com

2 Abdullah Nig. J. Fish. Aqua. 6(1) May, 2018 research is to investigate effect of honey on sex reversal and growth on Nile Tilapia (Oreochromis niloticus). MATERIALS AND METHODS This research was conducted at the Wet Laboratory of Kastela Faculty of Fishery and Marine Science, Khairun University, Ternate. Experimental fish Fish used in this research were 7-day old Oreochromis niloticus fry with an average length of 44 mm and average weight of g. The larvae were produced by natural spawning in the controlled tank at Fitu Hatchery facilities. Preparation of honey solution Honey used for the study was obtained locally from the Fuweu village market, Sula Regency, Indonesia. Different honey concentration (0 as a control, 1.0, 1.5 and 2.0 %) per litre were constituted in 2 litres of water. The honey and water were thoroughly mixed by stirring until the homogeneous solution was obtained. Experimental design Two hundred and forty (240) fry was used for the experiment. Different honey concentrations (1.0, 1.5, 2.0 and 0.0%) as a control, as treatments 1, 2, 3, and 4, respectively per litre of water were used for the immersion for 24 hours in a glass aquarium (40x40x40cm). Each treatment was replicated three times in randomized block design (CRD) manner. The fry was put in 2 litres of water that has been blended with honey in accordance with the concentration of each treatment. Twenty (20) fry were immersed in honey solution and left for 24 hours. Observation Phase The Nile Tilapia fry were reared for 10 weeks in which the sex of fish could be manually counted. During the culture period, water volume in the aquariums were added up to 20 litres as from day 30. RESULTS AND DISCUSSIONS Sex ratio Fry immersed in 2.0% honey gave the highest number of males. The result of the variance analysis showed that the treatment with the dosage of honey could produce the male sex that was significantly different compared to the control one. Meanwhile, in the treatment given honey did not give any significant influence from one treatment to others at the confidence level of 99%. From the result of the highest (86%) of male in all treatments was found in the treatment with the dosage of 2.0%, followed by treatment with the dosage of 1.5% with 84% males, treatment with the dosage of 1% at 77%, and the lowest one was found in treatment control with 38% male. The high number of males could be attributed to the active substance, chrysene and potassium at found the honey (Abdullah et al., 2015). Marhiyanto (1999) explained that each 100 gram of honey contains potassium in the range of ppm. IJEACCM (2006) as cited by Adhita et al. (2013) reported that honey contains compound known as chrysene that acts as a natural aromatase inhibitor. Syaifuddin (2004) stated that potassium can change the fat into pregnenolone that is known to be able to change the oestrogen into progesterone. Similarly, Syaifuddin (2004) and Saelistyawati et al, (2007) as cited by Ukhroy, (2008) also reported the high content of potassium in honey fed to also reported the high content of potassium in honey fed to GIFT fry might cause change in cholesterol located in the body tissue of the larvae into pregnenolone. Pregnenolone is the source of biosynthesis of steroid hormones (testosterone) by adrenal gland. 2

3 Abdallah Nig. J. Fish. Aqua. 5(1) May, 2017 Male (%) A(1%) B(1,5%) C(2,0%) D (0%) Hormone concetrations (%) Figure 1. Sex ratio of O. niloticus treated with honey. Given the change of oestrogen into progesterone, then the fish that is previously female could be reversed into the male. Aromatase inhibitor works by inhibiting the activities of aromatase. Such inhibition has led to the deterioration of concentration of oestrogen that later leads to deactivation of transcription from aromatase as the form of its feedback (Sever et al., 1999). The deterioration of the hormones of testosterone then turns to sex into the male. Work mechanism of aromatase inhibitor is to compete with the natural substrate of enzyme and interacts with the active side of enzymes, bind it and never return making the enzyme deactivated. However, the absorption of the chrysene by fish body is still relatively unknown which is one of the problems of chrysene regarding the chrysene absorption by the bloodstream Dean (2002) as cited by Utomo-Ukhroy, 2008). This achievement (87% males) in this study is assumed that the age of larvae (7 days) still in the phase of differentiation. As stated by Dunham (2004), the time of sex differentiation in Nile Tilapia occurs when the larvae are 6-7 days old after hatching up to around days old after hatching. Furthermore, Yamazaki (1983); Hunter and Donaldson (1983 in Muslim, 2011) explained that the artificial change in sex is possible as at the growing phase of gonad there is no any sex differentiation and the formation of steroid. Thus, the formation of gonad could be directed by using the synthetic steroid hormones. In addition, the environment becomes an influential factor particularly regarding the water temperature for rearing. It was found that the temperature is an environmental factor that plays an essential role towards the sex of fish (Strussman and Patino, 1995). The range of the temperature during the research was at C. In line with the statement of Baroiller et al. (1995), Wassel and Horstgen-Schwark (2007) as cited by Sarida et al. (2010) that temperature is an external factor that can bring an influence towards the change of female phenotype into the male phenotype in fish that have female phenotype. Histological examination of the sex of Nile Tilapia Figure 2 shows the male and female gonads of O. niloticus treated with 2% honey concentration. From the result of the observation using electron microscope, male gonad was characterized with the very small and dense formation of cell. In contrast, the female gonad was characterized with the larger cell with less dense among cells. 3

4 Abdullah Nig. J. Fish. Aqua. 6(1) May, 2018 Sperm Cell Embryo Egg Cell Embryo Figure 2: Male and female gonads of O. niloticus Treated with 2 % Honey Concentration Survival rate Survival rates (SR) in each treatment were at 57% -73% as shown in Figure 3 above. From the variance analysis, it was found that the highest level of survival rate was achieved in treatment B at 73%, respectively followed by treatment A and C at 72% and treatment D at 57%. The treatment given honey (treatment A, B and C) did not show any effect to each other. Once conducting the variance analysis, it was found that the treatment of the different immersion of the honey dosage within 24 hours showed a significant difference without any honey dosage (control). The treatments A (1.0%), B (1.5%) and C (2.0%) given with the honey dosage were significantly different from the treatment D without any honey dosage given (0%). It is in line with the result of variance analysis (F count 22 > F table=7.59). The immersion of 7-day larvae with the honey dosage in treatment A, B and C in fact had a significant influence on the treatment D control (without any honey immersion). This is assumed that honey has a content of nutrition that can give an immune for Nile larvae towards any disease and environment. It could be clearly seen at week 2 and 3 in which the larvae in treatment D were died in contrast to the treatment A, B and C given honey that were not died. Survival rate (%) 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% A B C D Honey concentration (%) Figure 3. Survival of Oreochromis niloticus treated with honey Survival rates in each treatment were at 57% -73% as shown in Figure 3 above. From the variance analysis, it was found that the highest level of survival rate was achieved in treatment B at 73%, respectively followed by treatment A and C at 72% and treatment D at 57%. The treatment given honey (treatment A, B and C) did not show any effect to each other. Once conducting the variance analysis, it was found that the treatment of the different immersion of the honey dosage within 24 hours showed a significant difference without any honey dosage (control). The treatments A (1.0%), B (1.5%) and C (2.0%) given with the honey dosage were significantly different from the treatment D without any honey 4

5 Abdallah Nig. J. Fish. Aqua. 5(1) May, 2017 dosage given (0%). It is in line with the result of variance analysis (F count 22 > F table=7.59). The immersion of 7-day larvae with the honey dosage in treatment A, B and C in fact had a significant influence on the treatment D control (without any honey immersion). This is assumed that honey has a content of nutrition that can give an immune for Nile larvae towards any disease and environment. It could be clearly seen at week 2 and 3 in which the larvae in treatment D were died in contrast to the treatment A, B and C given honey that were not died. Absolute biomass growth (W) Based on the result of the research conducted in 10 weeks towards the Nile Tilapia larvae reared in the aquarium with the treatment of immersion of honey dosage, it resulted in growth of average weight as shown in Table 4. Table 4. Absolute biomass growth in each treatment tested Honey Dosage (%) Absolute Biomass Growth (g) (Control) 0.95 Remark: the numbers given the equal letter in the same line and column were not significantly different from the confidence level of 99% (p>0.01). The tested fish reared in this research was the Nile tilapia with the initial average weight at week 0 was 0.01 g/fish up to week 10 with the average weight up to 1.05 g/fish from all treatment and has been categorized as seeds. Based on the variance analysis, it is found that the different treatment of immersion of honey dosage within 24 hours was not significantly different from one treatment to each other. (F count=3,82 F table=7,59). Figure 4 shows the average growth in each week. Water quality parameters The water quality during the rearing in this research was still in the tolerance limit for fish both in growth and survival. The average result in the measurement of water quality during the research can be seen in Table 5 below. Table 5. Mean value of water quality during research Honey concentration (%) Absolute Biomass Growth (g) (Control) 0.95 Conclusion and recommendation Honey at a dosage of 2.0% was highly effective in reversing sex in O. niloticus within 24 hours. The survival rate of in all the honey concentration were at 57% -73%. The growth was not significantly different from others. Based on this research, there is a need to conduct a further research on the by immersion at larval stage of sex differentiation not only to optimize the process of masculinization but also to investigate the growth and survival of the fish. REFERENCES Adhita A. D., Wayang S., and Wildan, (2013). Aplikasi Madu untuk Pengarahan Jenis Kelamin pada Ikan Nila (Oreochromis nilaticus). Journalof Aquaculture Program, Mataram University. Accessed in 10 May (7): Afonso, L.O., Wassermann, G.J. and Terezinha de Oliveira, R (2001). Sex reversal in Nile tilapia (Oreochromis niloticus) using a nonsteroidal aromatase inhibitor. Journal of Experimental Zoology, 290(2):

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