Are We There Yet?Zebrafish Nutrition

Stephen A. Watts, Louis D’ Abramo, Susan Farmer, Lacey Dennis, Daniel Smith and Mickie L. PowellUniversity 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

• Ingredients are practical

• Possible anti-nutritional 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

MethodsFish 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 25.00fish protein hydrolysate 20.00wheat starch 9.60wheat gluten 7.00alginate 5.38soy protein isolate 5.00dextrin 5.00menhaden fish oil 4.67soy lecithin (refined) 4.00vitamin premix 4.00mineral premix 3.00corn oil 2.33canthaxanthin (10%) 2.31potassium phosphate monobasic 1.15alpha cellulose 1.00glucosamine 0.25betaine 0.15cholesterol 0.12ascorbylpalmitate 0.04

Total 100.00

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

High BCILow

BCI

Can one nutrient affect experimental

outcomes?

ProteinMixed source vs sole source

Amino Acid

Analysis

MIXED FPI CAS SOY WGCysteine 0.400 0.400 0.190 0.530 1.010Methionine 1.080 1.160 1.320 0.560 0.730Lysine 2.930 3.200 3.610 2.700 0.660Alanine 1.894 2.915 1.427 1.932 1.249Arginine 2.090 2.748 1.642 3.187 1.547Aspartic Acid 3.284 3.849 3.322 4.996 1.509Glutamic Acid 9.631 5.626 10.120 8.388 18.060Glycine 2.072 3.984 0.876 1.848 1.622Isoleucine 2.023 1.709 2.394 2.132 1.763Leucine 3.631 2.866 4.354 3.500 3.293Serine 2.140 1.804 2.461 2.103 2.177Threonine 1.706 1.744 1.917 1.612 1.217Valine 2.424 2.019 3.016 2.196 1.868Histidine 1.120 0.940 1.337 1.110 0.975Phenylalanine 2.130 1.573 2.387 2.325 2.600Tyrosine 1.877 1.574 2.372 1.473 1.491Taurine 0.098 0.367 0.010 0.010 0.010Tryptophan 0.506 0.374 0.620 0.582 0.467

µCT of Mixed Protein and Soy Protein Diets

Bone Alterations

ConclusionsAll 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?

AcknowledgmentsUAB NORC Aquatic Animals Research

Core (NIH P30DK056336).

Jeff Barry, Adele Fowler, Chris Taylor, Michael Williams, Karen Jensen, Marlee Hayes

ACLAM