Are We There Yet?
Zebrafish Nutrition
Stephen A. Watts, Louis D’ Abramo,
Susan Farmer, Lacey Dennis, Daniel Smith
and Mickie L. Powell
University of Alabama at Birmingham and
Mississippi State University
Why Nutrition?
 ZF have been used extensively as a
drug and disease model
 ZF are a useful toxicology model
 Despite their widespread use the
nutritional requirements have not
been determined
AquacultureGrow large
fish fast.
AquaristsColor,
Reproduction
ResearchGrowth,
Overall
health, High
fecundity.
• Ingredients are
practical
• Possible antinutritional factors
• No indication of
quantities
• Use of lakes and
dyes
• Use of
preservatives
There are no standards among
diets in zebrafish studies!!
Where do we start?
Examples of Nutrition Studies
Considerations in the Use of Formulated Diets
 Age: Larval vs juvenile vs adult (breeding)
 Physical form of the diet
 Feed Frequency
 Protein source/quality
 Carbohydrate
 Lipid and Fatty Acids
 Gut Retention Time
 Bacteria
 Culture Container
Methods
 Fish were co-cultured with enriched
rotifers for the first 5 days post hatch
and fed Artemia for 11 days prior to
stocking.
 At 21 days post hatch 15 fish were
stocked randomly into 2.8 liter tanks
and maintained on a recirculating
zebrafish system.
 For each diet, fish were fed a ration in
excess of 5% of their body weight per
day, divided into two feedings.
 Fish were photographed and weighed
every two weeks to measure growth and
adjust feed rations.
UAB Z-12
Diet
Formulation
Ingredient
%
casein - vitafree
25.00
fish protein hydrolysate
20.00
wheat starch
9.60
wheat gluten
7.00
alginate
5.38
soy protein isolate
5.00
dextrin
5.00
menhaden fish oil
4.67
soy lecithin (refined)
4.00
vitamin premix
4.00
mineral premix
3.00
corn oil
2.33
canthaxanthin (10%)
2.31
potassium phosphate monobasic
1.15
alpha cellulose
1.00
glucosamine
0.25
betaine
0.15
cholesterol
0.12
ascorbylpalmitate
0.04
Total 100.00
Standard Growth Rate (SGR)
25
20
Z12
Zeigler
Tetramin
Gemma
Otohime
Artemia
SGR (%)
15
10
5
0
2
4
6
8
Weeks
10
12
Weights
900
Calculated Weight/Fish (mg)
800
Z12
Zeigler
Tetramin
Gemma
Otohime
Artemia
700
600
500
400
300
200
100
0
0
2
4
6
Week
8
10
12
Lengths
43
Average Length (mm)
38
33
28
Z12
Zeigler
Tetramin
Gemma
Otohime
Artemia
23
18
13
8
0
2
4
6
Week
8
10
12
Week 10
Artemia
Gemma
Diet
Body Condition Index
Artemia
0.98
Gemma
1.09
Z-12
1.19
Ziegler
1.24
Otohime
1.24
Tetramin
1.32
Low
BCI
High
BCI
Can one nutrient
affect experimental
outcomes?
Protein
 Mixed source vs sole source
Amino
Acid
Analysis
MIXED
Cysteine
0.400
Methionine
1.080
Lysine
2.930
Alanine
1.894
Arginine
2.090
Aspartic
Acid
3.284
Glutamic
Acid
9.631
Glycine
2.072
Isoleucine
2.023
Leucine
3.631
Serine
2.140
Threonine
1.706
Valine
2.424
Histidine
1.120
Phenylalani
ne
2.130
Tyrosine
1.877
Taurine
0.098
Tryptophan
0.506
FPI
0.400
1.160
3.200
2.915
2.748
CAS
0.190
1.320
3.610
1.427
1.642
SOY
0.530
0.560
2.700
1.932
3.187
WG
1.010
0.730
0.660
1.249
1.547
3.849
3.322
4.996
1.509
5.626 10.120
3.984
0.876
1.709
2.394
2.866
4.354
1.804
2.461
1.744
1.917
2.019
3.016
0.940
1.337
8.388 18.060
1.848
1.622
2.132
1.763
3.500
3.293
2.103
2.177
1.612
1.217
2.196
1.868
1.110
0.975
1.573
1.574
0.367
0.374
2.325
1.473
0.010
0.582
2.387
2.372
0.010
0.620
2.600
1.491
0.010
0.467
µCT of Mixed Protein and Soy
Protein Diets
Bone Alterations
Conclusions
 All diets supported growth and survival.
However, commercial diets contain undefined
ingredients.
 The consequence: Nutrients and nutrient source
affect outcomes.
 Any health or disease-related outcomes will be
affected by diet.
 Interpretation of experimental results must be
made within the context of a defined nutritional
history, or lack thereof.
 Microbiome?
Acknowledgments
 UAB NORC Aquatic Animals Research Core
(NIH P30DK056336).
 Jeff Barry, Adele Fowler, Chris Taylor,
Michael Williams, Karen Jensen, Marlee
Hayes
 ACLAM