OREOCHROMIS NILOTICUS

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1 OXYTETRACYCLINE MARKING STUDIES OF TILAPIA; OREOCHROMIS NILOTICUS Y. M. Abdel-Hadi 1, 2, J. F. Craig 3, J. A. Babaluk 4 and R. Wassle 1 Aquaculture Dept., Faculty of Agriculture, University of Putra Malaysia (UPM), Serdang, Selangor, Malaysia. 2 Central Laboratory for Aquaculture Research (CLAR), Abbassa, Egypt. 3 Whiteside, Dunscore, Dumfries, DG2 0UU, Scotland, U.K. 4 Freshwater Institute, 501 University Crescent, Winnipeg, Manitoba, Canada

2 I- Introduction Since oxytetracycline is incorporated into newly forming calcified tissue and is visible under ultraviolet light as a fluorescent band, it had been used periodically by fisheries researchers as a marking tool. Since that time, tetracycline has been used widely as a means of marking. Applied at a known date, tetracycline has been used to validate methods of fish age determination.

3 Importance of fish aging Fish age determination has been used in fisheries management: Stock assessment. Age of capture and marketing. Age of sexual maturity. Different Biological studies.

4 von Bertalanffy growth equation The individual growth model published by Karl Ludwig von Bertalanffy in 1934 is widely used in biological models and exists in a number of permutations. In its simplest version the so-called von Bertalanffy growth equation is expressed as a differential equation of length (L) over time (t): L/(t) = rb (L - L (t)) when r B is the von Bertalanffy growth rate, L is the ultimate length of the individual and t is the time or age.

5 II- Objectives In this study, tetracycline was injected into tilapia, Oreochromis niloticus to validate 4 age determination methods (scales, opercula, otoliths' and pelvic fin rays' sections): Tilapia fish aging reliability in Egypt. To assess tetracycline-induced mortality. To determine how quickly it was incorporated into calcifying tissues of Tilapia. To determine the suitable marking dosage of tetracycline for this freshwater species.

6 Materials and methods Fish: A number of 1200 fingerlings of monosex Oreochromis niloticus were used for the study. The fish were acclimatized first and distributed into 4 raceways (300 per each raceway). The average body weight at the time of injection of the OTC on September 1 st, 1998 was 60 gram and the average total length was 15 cm.

7 Raceways: Four raceways were used for rearing the fish for an entire year, which was the period of the experiment. The dimensions of each raceway were 50 meters long, 4 meters width and 1 meter of water depth. The experiment was carried out in the facilities of the WorldFish Centre (The Regional Centre for Africa & west Asia at Abbassa,, Egypt) for 1 year.

8 Selection of Oxytetracycline dosages The fish in the 1 st group were injected with OTC in a dose of 12.5 mg/kg fish body weight and then released into the first raceway. The fish in the 2 nd group were injected with OTC in a dose of 25 mg/kg fish body weight and then released into the second raceway. While, the fish in the 3 rd group were injected with OTC in a dose of 50 mg/kg fish body weight and then released into the third raceway.

9 On the other hand, the fish of the 4 th group were injected with sterile distilled water and then released to the 4 th raceway to be kept as a control group. The fish were injected intraperitoneally (I/P) using an automatic syringe similar to that used in mass injection of chickens in poultry farms.

10 Automatice Syringe

11 Sampling regime One fish from the fish kept in the control hapa and 5 from the fish injected with 50 mg/kg.b.wt. were taken periodically after 6, 12, 24, 48, 72 and 96 hours. Then after 1, 2 and 4 weeks respectively. Periodical sampling of 5 fish from the 4 treatments in a monthly basis was carried using trammel net.

12 Those 5 fish from each treatment were sacrificed. Each fish was numbered, weighed and the total body length was measured. The fish were then dissected. Different bony structures were taken from each fish, which are scales, opercula, otoliths and pelvic fins.

13 Examination of different bony structures Opercula: were examined whole on a black background with a stereoscopic microscope with a handheld reflective ultraviolet light source. The viewing room was dark. Scales, otolith sections, and fin ray sections were examined under reflected ultraviolet light using a fluorescence microscope.

14 Age validation: Results There was quite a bit of growth on all structures after the OTC mark. It also appears that the fish were at large long enough so that an annulus or check had been formed after the OTC mark. This was especially evident on the pelvic fin rays and opercula. Thus, all examined bony structures were found to be reliable and valid for aging of Oreochromis niloticus in Egypt.

15 Time of annulus formation was on May depending on scales' back calculations (Figure. 1). On the other hand, its time may be on April according to opercula's back-calculations (Figure. 2).

16 Length in mm Fig. 1 The relationship between age, total body length (T.L.) & scale radius (R) Date Average T.L. mm R mm 0 Date from September 1998 to September 1999 Figure. 1 Back-calculations using scales radii N.B. Notice the stunted growth during winter months (In January, February & March). R = Average total scale radius in mm (sampled from 5 fish).

17 Length in mm 300 Fig.2 The relationship between age of fish, total body length and operculum radius Date in month Total body length in mm total operculum radius (R in mm) Date from September 1998 to September 1999 Figure. 2 Back-calculations using opercula radii R = Average total operculum radius in mm.

18 Oxytetracycline-induced mortality Treatment Total mortality 12.5 mg/kg fish 2.7% 25 mg/kg 0.3% 50 mg/kg 1.3% Control group 2.7 %

19 Oxytetracycline uptake & dosage All the 3 dosages were sufficient to mark the various bony structures. OTC marks were clearly evident on all structures from OTCinjected fish that were at large for one month. The 25 and 50 mg/kg gave strong OTC marks In all of these cases, there was visible growth on the bony structure after the OTC mark.

20 Figure. 3 Transverse section of an otolith from a tilapia injected with 25 mg oxytetracycline/kg body weight (a) reflected ultraviolet light and b) transmitted white light. Annuli are indicated by dots. Oxytetracycline mark (OTC) is also indicated. Bar = 0.5 mm.

21 Figure. 4 Transverse section of a pelvic fin ray from a tilapia 9 months after oxytetracycline injection

22 Figure. 5 Transverse section of an otolith from a tilapia 9 months after oxytetracycline injection

23 Figure. 6 Transverse section of an otolith from a tilapia 2 months after oxytetracycline injection

24 Figure. 7 Transverse section of an otolith from a tilapia 48 hours after oxytetracycline injection

25 Conclusion It could be concluded that tilapia fish aging is reliable in Egypt as a subtropical country and can be used as a basis for stock assessment and other fisheries management applications. The dose of 25 mg OTC/kg body weight for marking of tilapia is recommended in subsequent work & future studies.

26 Acknowledgment The authors are very grateful to the WorldFish Centre and its entire staff (Regional Center for Africa and West Asia, Abbassa, Egypt), where the experiment was carried out. Special thanks to Dr. Roger Row (the former deputy director general), Engineer Rezk Hara and his team. The authors also appreciate the kind support of Dr. Ismail Radwan who granted the fish of the experiment as a gift from his own hatchery in Kafr El-Sheikh, Egypt. Sincere thanks are directed to UPM for sponsoring my travel and participation in this big event.

27

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