The aim of this study was to improve the rearing of common clownfish Amphiprion percula, by weaning juveniles from a live feed to a formulated feed as early as possible, while still maintaining good growth and survival. The growth response of A. percula to a formulated dry feed was initially investigated. There was no difference in growth rate of juveniles fed a formulated feed, and the formulated feed supplemented with either Artemia or a Donax serra / Penaeus indicus combination. Amphiprion percula readily consumed the formulated feed, and the fishmeal/casein combination appeared an acceptable protein source. As the amount of protein included in a diet can have a profound effect on growth, the optimal dietary protein level for juvenile A. percula was investigated by feeding semipurified diets containing graded levels of protein, ranging from 40-65%. There was no difference in the growth rates of juveniles fed the various diets, however all diets promoted good growth with an average weight gain of 419%, and thus for the purposes of this study the diet formulation was deemed adequate. The histological study of the digestive system of larval A. percula revealed that the alimentary canal was advanced at hatching and that larvae start exogenous feeding immediately. Three days after hatch (DAH) the yolk sac is completely absorbed. In the hind-gut epithelium of 5-day-old larvae small supranuclear inclusion vacuoles appear, suggesting pinocytotic digestion, and by 7 DAH gastric glands are established in the epithelium of the stomach. Nine DAH supranuclear inclusion vacuoles appear in the epithelium of the mid-gut, indicating extracellular digestion and absorption across the lumen. As pinocytotic digestion of protein is less efficient than extracellular digestion, especially in the case of formulated feeds, it was hypothesised that the digestive system of A. percula could only effectively digest formulated feeds 9 DAH onwards. The two weaning experiments designed to test this hypothesis revealed that A. percula was able to utilise the formulated feed, without reduction in survival, from 7 DAH onwards. However, in terms of growth, the optimal time to wean juveniles from the live feed to the formulated dry feed was between 15 to 20 DAH. As A. percula accept a formulated feed and can benefit nutritionally from it, the dependence of larvae and juveniles on live feed can be reduced. This study has shown that the rearing of A. percula can been simplified and improved by weaning from 7 DAH with no reduction in survival, and from 15 to 20 DAH with no reduction in growth.

Thesis--Auburn University, 2007. / Abstract. Includes bibliographic references (p.117-142).

Thesis (Ph.D.) - James Cook University, 2005. / Typescript (photocopy) Bibliography: leaves 143-164.

In aquaculture, a thorough knowledge of the specific environmental requirements of a species is needed in order to maximize growth rate and survival. There is a paucity of data regarding the fundamental environmental requirements for the ongrowing phase of clownfish juveniles. This prompted the design of three experiments to determine the best feeding regimen, temperature and stocking density that maximize growth and survival of Amphiprion percula. Ration size and feeding frequency are important factors for optimizing fish growth during the juvenile grow-out phase. A factorial growth trial was conducted to determine the effect of feeding frequency and ration size on the growth of juvenile clownfish (Amphiprion percula).Three feeding frequencies (1, 2 and 3 times daily) and six rations (2,4,6,8, 10 and 12 % body weight per day (BW.day⁻¹)) were used to test the growth response over a twelve week period. Non-linear regression analysis on the effect of ration, independent of feeding frequency, on growth resulted in a significant (n = 36; r² = 68.7) parabolic model: In y = -0.0302x² + 0.5159x + -4.4377. Maximum growth corresponded to a ration of 8.5% BW.day⁻¹. Survival as a function of ration was significantly lower at 2% BW.day⁻¹. Data were further examined with Analysis of CoVariance (ANCOVA) to determine the effect of ration on growth at each feeding frequency. The combination revealed a maximum growth rate when the fish were fed a ration of 10% BW.day⁻¹ divided into two equal meals. The required ration per meal to maintain maximum growth was also found to decrease as feeding frequency increased. The determination of the best temperature for growth is of great importance due to the direct relationship between fish metabolism and temperature. The thermal preferendum of A. percula has already been estimated at 26 ± 0.7°C but it is not known whether this closely approximates the temperature for maximum growth. Sixteen tanks were set to different temperatures ranging between 21.5 and 30.2 °C. Ten juvenile A. percula were placed in each tank and growth was measured fortnightly over the course of the 10 week experiment. Non-linear regression analysis resulted in significant models for fish length (y = -0.0005x² + 0.00267x - 0.0338; r² = 56.7, n = 11) and weight (y= -0.00016x² + 0.0084x - 0.1073; r² = 61.6, n = 11). These models predict that maximum growth would be at 27.7 and 27.1 °C for length and weight, respectively. Temperature, over the range tested, had no apparent effect on survival. There was no significant difference between the temperatures for maximum growth and the preferred temperature. The effect of stocking densities ranging from 0.2 fish.L¹ to 4.0 fish.L⁻¹ on growth were used in the third experiment. The fish were fed to satiation twice daily and growth was measured fortnightly throughout the 8 week experiment. No effects on growth, survival or coefficient of variation were found within the range of stocking densities tested.

The objectives of the study were to determine whether or not behaviour could be used to predict spawning in Amphiprion akallopisos, and to document the behaviour of this species under various environmental conditions in captivity. The spawning behaviour of A. akallopisos was studied and quantified. Three behaviour patterns - belly touching, nest cleaning by the female and mutual nest cleaning (by the male and the female) were identified as predictors for spawning. The reproductive behaviour of A.akallopisos under three photoperiods was investigated. The photoperiods were: 14L:10D, 10hr15minL:13hr45minD and a natural photoperiod cycle condensed into three months. A. akallopisos maintained under 14 L:10D exhibited a significantly higher frequency and duration of chasing, nest cleaning and total interaction compared to fish kept under 10hr15minL:13hr45minD. A photoperiod of 14L:10D was selected for further studies. As manipulation of photoperiod did not induce spawning, GnRHa was administered to the fish in the diet at levels of 10, 20, 40 and 80 μg/kg BW. Control groups were fed untreated food. None of the dosages were successful in inducing spawning, or spawning behaviour in A. akallopisos. It is possible that the method of hormone application was not suitable for this species. It is also possible that behaviour may regulate blood hormone levels as opposed to endocrine status influencing behaviour. In the third trial, A. akallopisos was maintained with, and without sea anemones in order to determine whether or not anemone hosts are necessary for spawning. The presence of anemones did not induce spawning and A. akallopisos kept without anemones exhibited significantly more interactive behaviour than fish kept with hosts. Spawning of A. akallopisos at a later date without anemones suggests that anemones are not necessary for spawning in A. akallopisos. Light intensity was investigated as a cue for spawning. The reproductive behaviour of A. akallopisos was then studied under light intensities of 4.16 x 10¹⁵ quanta.sec⁻¹.cm⁻², 8.85 x 10¹⁵ quanta.sec⁻¹.cm⁻² and this intensity plus natural light. Spawning occurred mainly under 8.85 x 10¹⁵ quanta.sec⁻¹.cm⁻². Fish maintained under this light intensity exhibited significantly more of nest cleaning behaviour in terms of frequency and duration than fish maintained under low light intensity. A minimum light intensity of 8.85 x 10¹⁵ quanta. sec⁻¹.cm⁻² is recommended for conditioning of this species. This study can be of practical relevance to hatchery managers who can use the methods developed and record predictors for spawning in A. akallopisos and other Amphiprion species. In this way imminence of spawning may be estimated. Most importantly, the study also has academic merit as little work has been undertaken in this field. Although the observational method used in this study was adequate for the purposes of the investigation, future work of this nature should incorporate other methods of documenting gonadal development such as gonadal staging and GSI in order to obtain more conclusive results.