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Saponin fractions from fenugreek (Trigonella foenum-graecum L.)as dietary supplements for Nile tilapia (Oreochromis niloticus L.)and common carp (Cyprinus carpio L.)
(2012) Stadtlander, Timo; Focken, Ulfert
The future role and importance of aquaculture for the world food supply is more and more recognized by the public. High quality feed for semi-intensive and intensive aquaculture operations are necessary to guarantee a future continuous growth of fish production. Nutrient utilization efficiency has in the past been augmented by addition of feed additives, mainly anti-biotics and hormones. That practice is prohibited in the European Union since 1st of January 2006 and the consumer acceptance of products produced in that way is low. Around a decade ago saponins, secondary plant metabolites, have been found to be potential alternatives for aquafeeds. Saponinmixtures derived from the South American soap bark tree Quillaja saponaria, have been found to have beneficial effects on growth, feed and nutrient utilization and on metabolism of common carp Cyprinus carpio and Nile tilapia Oreochromis niloticus. Furthermore, in one of the early experiments a sex ratio in favor of males was observed in saponin fed tilapia compared to control fish not supplemented with saponins. An influence on sex ratio of Nile tilapia would be highly desirable since industrially Nile tilapia production is only financially viable if all male populations are produced since mixed sex tilapia populations tend to reproduce uncontrollable. The production of all male tilapia populations is mainly conducted by feeding the potentially environmentally hazardous and carcinogenic synthetic hormone 17--methyltestosterone to sexually undeveloped tilapia fry. The above mentioned problems led to a joint research project with partners in Israel and Palestine in which this dissertation has been conducted. The aim of the project was the fractionation of crude saponin extract derived from fenugreek Trigonella foenum-graecum and its testing in vitro and in vivo. It was assumed that a higher biological activity could be achieved if saponin fractions instead of crude mixtures would be applied. After in vitro testing, different saponin fractions or eluates, have been tested on their effects on growth, feed and nutrient utilization, metabolism, proximate composition, gene expression of GH and IGF-1, sex ratio, reproduction and gonad histology. Most of the feeding experiments were conducted with Nile tilapia but one experiment was also conducted with carp. Three of the feeding experiments have been conducted in a system capable of measuring continuous respiration of the fish. Another experiment has been conducted at a field station at Jericho, Palestine. The experiments conducted to evaluate the influence of saponin fractions on sex ratio of undifferentiated tilapia fry have been conducted primarily in a flow-through system. The tested saponin fractions and a tested sapogenin are not suitable to produce male monosex tilapia populations. The sex ratios after supplementation of diets of mixed sexed tilapia fry did only in one case show a significantly higher proportion of males than the control. In a larger scale repetition of that experiment the previous observations could statistically not be proofed. It must be assumed to be random effects or be the result of initially biased sex ratios after stocking undifferentiated tilapia fry. An experiment in which genetically female tilapias were fed with two fenugreek saponin fractions and positive and negative controls supported that finding. A long term feeding experiment conducted at Jericho revealed no influence of the long and short term supplemented Q. saponaria saponin mixtures on sex ratio and reproduction of genetically female tilapia. A similar laboratory experiment with fenugreek saponin fractions could not be evaluated in that regard. The experiments evaluating the effects of the saponin fractions on growth, feed and nutrient utilization, gene expression of GH and IGF-1 and proximate composition revealed no significant differences. But in all experiments one fraction eluated with 60%/40% (v/v) methanol/water showed numerically improved values compared to control and other fractions. Furthermore one fraction eluated with 40%/60%/ (v/v) methanol/water gave constantly numerically inferior results of tested parameters compared to control. That supports the conclusion that the 40% methanol fraction contains saponins generally referred to as anti-nutrients. In only one experiment, conducted at Jericho, significantly higher growth was observed after long term supplementation with a Q. saponaria saponin mixture containing elevated sapogenin content. The presented data in this thesis is not supporting an application of the tested saponin fractions as environmentally friendly alternative to methyltestosterone to produce all male populations of tilapia or to inhibit uncontrollable reproduction. Additional experiments are needed to evaluate different modes of application like immersion treatments or injections since during the experimental work of this thesis all tested saponins were added to the feed. An application of the tested fenugreek saponin fractions as growth promoters yields a higher potential although the experimental results are based upon low sample sizes due to capacity restrictions of the respirometric system. Therefore a repetition under near commercial or commercial conditions must be considered.