DIURNAL PATTERNS OF TRYPTIC ENZYME ACTIVITY IN SEA BREAM LARVAE UNDER DIFFERENT FEEDING REGIMES M.Sc. Sinem Zeytin Gesellschaft für Marine Aquakultur, Büsum Christian-Albrechts-Universität zu Kiel 6.Büsumer Fischtag 11.06.2015
Digestive processes in larval stages 2
Trypsin (hydrolised MCA, nmol min -1, larva -1 ) Daily rhythm of Tryptic enzyme activity of sardine larvae in nature Mean length Mean Tryptic activity Length (mm) Sampling Time (hours) (Ueberschär, 1995) 3
Phase II: critical larval stage with decreasing tryptic activities, poor growth and high mortality rates Phase III: sufficient production of trypsin (at optimal food supply), high growth rates Phase I: yolk-sac stage and beginning of exogenous feeding Phase IV: beginning of metamorphosis, during which tryptic activity is being partly replaced by the activity of pepsin in the (after Ueberschär, 2000) developing stomach 4
Trypsin is the most significant protease in the early larval stages and is considered to be a key enzyme in the digestive process This short-term experiment was conducted to evaluate the impact of different dietary treatments on the diurnal digestive tryptic enzyme activity in gilthead sea bream (Sparus aurata) larvae 5
Larval Rearing Facility Stocking density 75 larvae L -1 in 65 L rearing tanks 4 different age groups (21 & 26 & 34 & 44 dph) 24 hours illumination 20 hour short-term experiment (06:30 01:30) 4 different feeding regimes (live feed and MD) 6
26 & 44 dph 44 dph Every 15 min 30 mg/feeding event 4 Artemia ml -1 4 & 6 & 15 Rotifers ml -1 6 R.ml -1 +1.3 A.ml - 1 13.26 5.99 6.74 8.22 R & A phase Rotifer phase Commercial diet Artemia phase 21 & 26 & 34 dph 26 dph 7
C C C C n= 6 sea bream larvae sampled from each group in each hour The tryptic enzymatic activity of 1680 individual larva sample was analyzed Time Treatments Age Length Tryptic enzyme activity 8
Tryptic enzyme activity (hydrolised MCA, nmol min-1, larva-1) Results R21 Ø 7.05 mm R26 Ø 7.24 mm R34 Ø 8.11 mm All data points are the means of n=6 sea bream larvae 9
Tryptic enzyme activity (hydrolised MCA, nmol min-1, larva-1) Results R21 R26 R34 All data points are the means of n=6 sea bream larvae 10
Tryptic enzyme activity (hydrolised MCA, nmol min-1, larva-1) Results Ø 6.76 mm Ø 6.40 mm Ø 7.05 mm 20 18 16 14 12 10 8 6 4 2 0 RA 26 Fed with Rotifer + Artemia Fed with Microdiet MD 26 R 26 Fed with Rotifers Unfed larvae Sampling time All data points are the means of n=6 sea bream larvae 11
Tryptic enzyme activity (hydrolised MCA, nmol min-1, larva-1) Results Ø 6.76 mm Ø 6.40 mm Ø 7.05 mm 20 18 16 14 12 RA 26 MD 26 R 26 Mean T.A. of RA 26 Mean T.A. of MD Fed with Rotifer + Artemia Fed with Microdiet Fed with Rotifers Unfed larvae Mean T.A fed with Rotifer + Artemia Mean T.A. fed with MicroDiet 10 8 6 4 2 0 Sampling time All data points are the means of n=6 sea bream larvae 12
Tryptic enzyme activity (hydrolised MCA, nmol min-1, larva-1) Results MD 26 Sampling time All data points are the means of n=6 sea bream larvae 13
Tryptic enzyme activity (hydrolised MCA, nmol min-1, larva-1) Results Ø 12.6 mm Ø 13.9 mm 200 180 160 140 A 44 Fed with Artemia MD 44 Fed with MicroDiet Unfed larvae 120 100 80 60 40 20 0 Sampling time All data points are the means of n=6 sea bream larvae 14
Tryptic enzyme activity (hydrolised MCA, nmol min-1, larva-1) Results Ø 12.6 mm Ø 13.9 mm 200 Fed with Artemia 180 160 140 120 100 80 60 40 20 0 A 44 MD 44 Fed with MicroDiet Mean T.A. of A 44 Unfed larvae Mean T.A. fed with Artemia Mean T.A. of MD 44 Mean T.A. fed with MicroDiet Sampling time All data points are the means of n=6 sea bream larvae 15
Conclusions The results indicate that no matter what kind of diet was applied, there is a limited digestive capacity in sea bream larvae at some point in time during the day There is an existence of endogenous rhythm as internal biological clock in daily rhythm The results should be considered as speciesspecific development stage These findings should be considered in feeding schedules for younger larval stages 16
Outlook In order to investigate the digestive enzyme capacity, a comparison study should be done between continously and not continously fed larvae Other enzymes (Pepsin, Amylase, Lipaz) should be investigated in order to evaluate the digestive processes development 17
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