A Novel Ingredient for Zebrafish Diets: Single Cell Protein
Transcript of A Novel Ingredient for Zebrafish Diets: Single Cell Protein
![Page 1: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/1.jpg)
A Novel Ingredient for Zebrafish Diets: Single Cell Protein
Stephen A. Watts*, Robert J. Barry, Sophie Bru, Yuan Yuan,
L. Adele Fowler, Michael B. Williams,
Mickie L. Powell, Addison L. Lawrence
The University of Alabama at Birmingham
![Page 2: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/2.jpg)
ProteinPercent
Source
MineralsCalcium
Iron
Magnesium
Manganese
Selenium
Copper
CarbohydratesComplex
Simple
Source
LipidsSaturated
Unsaturated
Marine
Plant
VitaminsC, E, A, K, D
Thiamine
Niacin
Riboflavin
![Page 3: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/3.jpg)
Common Protein Sources in Aquafeeds
• Fish (type, meal, hydrolysate, isolate, etc.)• Casein• Soy• Squid Meal• Krill meal• Whey• Gluten• Gelatin• Feather and Poultry Meal• All vary in amino acid content and other bioactive nutrients
![Page 4: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/4.jpg)
Fish Meal in Animal Feeds
What is Fish Meal?
•Wild-caught, fish that have been dried
and ground to be used as a feed
ingredient
• Very digestible and palatable
• Balanced amino acid, phospholipid,
and fatty acid profiles
• Excellent source of protein
• Every zebrafish diet contains fish meal
![Page 5: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/5.jpg)
• A big problem: Global demand for fish meal protein has increased significantly yet production remains static
• Aquaculture industry accounts for ca. 60-70% of fish meal usage
• The product can be highly variable in quality
• A sustainable alternative to fish meal is necessary to meet the future demands of the aquaculture industry
Fish Meal Demand is Increasing
![Page 6: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/6.jpg)
So what about zebrafish diets?Should we be concerned about fish meal in zebrafish diets?
Should we be concerned about sustainability?
Should we be concerned about variability of the product?
Should we be concerned about reproducibility?
Alternatives?
![Page 7: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/7.jpg)
Fish Meal Alternatives
Source Examples Advantages Disadvantages
Plant Based Proteins Soybean mealCanola mealCorn gluten mealCottonseed meal
Generally high in protein content (40-70%) with the exception of wheat (~11%)
Deficient in one or more essential amino acids Anti-nutritional factors present in some ingredientsGrowth suppression
Animal Byproducts Feather mealPoultry byproducts Bloodmeal
High protein contentBlood meal rich in lysine
Most are deficient in lysine, methionine, and/or histidineBlood meal highly sensitive to heat damageLow in omega-3 & minerals
Trimmings Fish byproductsFish carcass meals
Best substitute for fish meal as it contains similar nutritional profiles
Unavailable in sufficient quantities to support fish meal replacement
![Page 8: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/8.jpg)
8
Single Cell Protein (SCP)• High nutrient content and quality
• Amino acid profile similar to FM
• Cheap
• Production can be controlled
• Environmentally sustainable
Nutrient composition of the main group of microorganisms used for single cell protein production(% dry wt.)
SCP- dried cells of microorganisms such as bacteria, microalgae, or yeast
Composition Algae Yeast Bacteria
Protein 40-60 45-55 50-65
Fat 7-20 2-6 1-3
Ash 8-10 5-10 3-7
Miller and Litsky 1976
![Page 9: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/9.jpg)
Question: Can SCP replace fish meal protein?• 35-day old zebrafish were fed a formulated diet containing
casein, soy, and fish protein hydrolysate (FPH) as the protein sources
• The fish protein hydrolysate was replaced with either a dried yeast or dried bacteria commercial product on a nitrogen basis
• Control formulated diets contained FPH; in experimental diets 50% or 100% of the FPH was replaced by the yeast or bacterial product
• Zebrafish were fed at either 10% body weight per day (satiation) or at 4% body weight per day (sub-satiation)
• All the usual environmental parameters were followed
![Page 10: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/10.jpg)
Water Quality
Mean Max Min
Temperature (°C) 27.7 ± 0.24 28.2 27.2
Conductivity (µs/cm) 1505± 48 1617 1425
Salinity (ppt) 0.71 ± 0.027 0.80 0.70
pH 7.67 ± 0.15 7.960 7.12
Ammonia (mg/L) 0.000 ± 0.000 0.000 0.000
Nitritie (mg/L) 0.000 ± 0.000 0.000 0.000
Nitrate (mg/L) 92.5 ± 10.35 100.0 80.0
![Page 11: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/11.jpg)
![Page 12: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/12.jpg)
Diet Ration Survival (%) ± SEM
FPH-100 10 100.0 ± 0.00
DY-50 10 100.0 ± 0.00
DY-100 10 100.0 ± 0.00
MRD-50 10 100.0 ± 0.00
MRD-100 10 100.0 ± 0.00
FPH-100 4 100.0 ± 0.00
DY-50 4 99.0 ± 1.01
DY-100 4 100.0 ± 0.00
MRD-50 4 99.0 ± 1.19
MRD-100 4 100.0 ± 0.00
RESULTS
![Page 13: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/13.jpg)
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0 2 4 6 8
Week
Ave
rage
Ind
ivid
ual
Wei
ght
(g)
Wet Weight (g)
FPH/100/10
DY/100/10
DY/50/10
MRD/100/10
MRD/50/10
FPH/100/4
DY/100/4
DY/50/4
MRD/100/4
MRD/50/4
![Page 14: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/14.jpg)
Diet Ration (% body weight) Male mg (SEM) Female mg (SEM)
FPH-100 10 249 (6.8)a 325 (11.6)a
DY-50 10 256 (8.7)a 328 (21.3)a
DY-100 10 275 (8.4)ab 379 (19.2)a
MRD-50 10 309 (6.8)b 341 (14.2)a
MRD-100 10 297 (7.8)b 323 (14.0)a
FPH-100 4 182 (3.6) 187 (7.5)
DY-50 4 185 (4.4) 186 (8.2)
DY-100 4 184 (4.9) 204 (8.6)
MRD-50 4 170 (4.0) 177 (6.5)
MRD-100 4 166 (4.6) 179 (8.0)
![Page 15: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/15.jpg)
Apparent Feed Conversion Ratio* (SEM)
Diet Ration Total FCR
FM-100 10 3.11 (0.380)
DY-50 10 3.61 (0.543)
DY-100 10 3.35 (0.976)
MRD-50 10 2.60 (0.358)
MRD-100 10 2.74 (0.317)
FM-100 4 1.34 (0.052)
DY-50 4 1.30 (0.057)
DY-100 4 1.25 (0.045)
MRD-50 4 1.40 (0.073)
MRD-100 4 1.46 (0.063)
*Grams feed proffered vs grams weigh gain
![Page 16: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/16.jpg)
Diet Ration Mean Moisture % (SD)
FPH-100 10 71.8 (0.83)a
DY-50 10 71.8 (1.16)a
DY-100 10 71.2 (0.81)a
MRD-50 10 72.1 (1.31)ab
MRD-100 10 72.3 (0.97)ab
FPH-100 4 74.5 (0.58)cd
DY-50 4 73.6 (0.42)bc
DY-100 4 73.5 (0.34)bc
MRD-50 4 75.5 (0.31)d
MRD-100 4 74.5 (0.76)cd
![Page 17: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/17.jpg)
BA
B
0
5
10
15
20
25
30
35
40
% L
ipid
Diet
Males
FPH/100/10 Male MRD/100/10 Male DY/100/10 Male
NSNS NS
0
5
10
15
20
25
30
35
40
% L
ipid
Diet
Females
FPH/100/10 Female MRD/100/10 Female DY/100/10 Female
![Page 18: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/18.jpg)
Conclusions
• Growth rates equaled or exceeded FPH controls when FPH is replaced with 50 or 100% yeast or bacteria SCP
• Male zebrafish fed bacteria SCP showed an increase in lean matter weight gain and reduced adiposity in relation to those fed FPH
• Both yeast and bacterial SCP show promise as fish protein replacements, but require additional health and reproductive evaluations
• We do NOT suggest that fish meal protein should be replaced in zebrafish diets. Fish meal has proven to be an excellent protein source in zebrafish “chows”. However, SCP may have importance in “formulated reference diets” where reproducible dietary control is valued
![Page 19: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/19.jpg)
Acknowledgements
19
❖ Meridian Biotech LLC.
❖ Members of the lab
❖ UAB Department of Biology
❖ Alabama Academy of Science
❖ NIH NORC P30DK056336
![Page 20: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/20.jpg)
Proximates FM-100 DY-50 DY-100 MRD-50 MRD-100Moisture % 12.73 13.43 12.84 14.87 16.64
Protein % 43.4 43.3 43.4 41.2 40.8
Fat % 11.1 9.95 10.3 12.5 9.11
Fiber % 0.8 2.4 1.6 1.3 nd
Ash % 6.66 6.24 5.96 7.57 8.94
TDN % 82.2 75.1 76.5 76.2 70
sulfur % 0.53 0.53 0.56 0.49 0.44
phosphorous % 1.32 1.32 1.4 1.72 2.17
potassium % 0.92 0.75 0.75 0.95 0.98
magnesium % 0.04 0.07 0.11 0.26 0.49
calcium % 0.83 0.81 0.94 0.78 0.78
sodium % 0.84 0.71 0.72 0.73 0.64
iron ppm 93.4 150 205 129 172
manganese ppm 98.3 101 112 104 113
copper ppm 7.6 16.6 30.7 9.8 13.7
zinc ppm 39.2 53 70 71.2 105
![Page 21: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/21.jpg)
• Funding: Meridian Biotech, LLC
NIH NORC: P30DK056336
• Special thanks to:Adele Fowler
Michael Williams
Chris Taylor
Audrey Powers
Ningxiang Zeng
Susmita Patnaik
Acknowledgements
![Page 22: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/22.jpg)
22
Single Cell Protein (SCP)
• High nutrients content and quality
• Amino acid profile similar to FM
• Cheap and Effective
• Selectable
• Environmentally sustainable
Nutrient composition of the main group of microorganisms used for single cell protein production(% dry wt.)
SCP- dried cells of microorganisms such as bacteria, microalgae, or yeast
Composition Fungi Algae Yeast Bacteria
Protein 30-45 40-60 45-55 50-65
Fat 2-8 7-20 2-6 1-3
Ash 9-14 8-10 5-10 3-7
Miller and Litsky 1976
![Page 23: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/23.jpg)
Amino Acid Fish Meal % Yeast SCP % Bacterial SCP %
Aspartic Acid 5.8 3.61 5.16
Threonine 2.7 2.46 1.99
Serine 2.5 2.80 1.90
Glutamic Acid 8.6 11.50 8.22
Arginine 4 2.19 2.57
Glycine 4.8 2.10 3.19
Alanine 4.2 4.44 5.55
Proline 3.3 5.69 2.37
Cystine 0.6 1.13 0.35
Valine 3.2 3.02 3.53
Methionine 2.2 1.14 1.17
Isoleucine 2.7 2.35 2.89
Leucine 4.6 8.56 4.47
Tyrosine 2.1 2.70 2.10
Phenylalanine 2.5 3.49 2.80
Histidine 1.8 1.48 0.98
Lysine 5 1.36 3.75
Tryptophan 0.5 0.41 0.62
Amino Acid content of SCPs (% dry wt.)
![Page 24: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/24.jpg)
• Single Cell Proteins (SCP) –dried cells of microorganisms such as yeast, bacteria, or microalgae
• Amino acid profile similar to fish meal
• Sustainable
Amino Acid Fish Meal % Yeast SCP % Bacterial SCP %
Cystine 0.6 1.13 0.35Valine 3.2 3.02 3.53Methionine 2.2 1.14 1.17Isoleucine 2.7 2.35 2.89Leucine 4.6 8.56 4.47Tyrosine 2.1 2.70 2.10Phenylalanine 2.5 3.49 2.80Histidine 1.8 1.48 0.98Lysine 5 1.36 3.75Arginine 4 2.19 2.57Tryptophan 0.5 0.41 0.62
Single Cell Proteins as Fish Meal Replacement
![Page 25: A Novel Ingredient for Zebrafish Diets: Single Cell Protein](https://reader031.fdocuments.net/reader031/viewer/2022012513/618bffa1cead60139156a060/html5/thumbnails/25.jpg)
Ingredient
Z17-0201
FM-100
Z17-0202
DY-50
Z17-0203
DY-100
Z17-0204
MRD-50
Z17-0205
MRD-100casein - low trace metals 20 20 20 20 20
soy protein isolate 6.6 6.6 6.6 6.6 6.6
fish protein hydrolysate 25 12.5 12.5
SCP Sample 1 (DY) 18.5 37
SCP Sample 2 (MRD) 15.37 30.75
wheat starch 15.95 11.48 7 12.08 8.2
dextrin 5.5 5.5 5.5 5.5 5.5
menhaden fish oil (ARBP) 0 1.25 2.5 1.25 2.5
corn oil 5.55 2.77 5.3 5.05
soy lecithin (refined) 4 4 4 4 4
alginate (TIC algin 400) 5.38 5.38 5.38 5.38 5.38
vitamin mix (MP-VDFM) 4 4 4 4 4
mineral mix (BTm) 3 3 3 3 3
canthaxanthin (10%) 2.31 2.31 2.31 2.31 2.31
potassium phosphate monobasic 1.15 1.15 1.15 1.15 1.15
alpha cellulose 1.14 1.14 1.14 1.14 1.14
glucosamine 0.11 0.11 0.11 0.11 0.11
betaine 0.15 0.15 0.15 0.15 0.15
cholesterol 0.12 0.12 0.12 0.12 0.12
ascorbylpalmitate 0.04 0.04 0.04 0.04 0.04
Total 100 100 100 100 100
Calculated Energy Level (kcal) 3780 3600 3422 3624 3469
Calculated Protein Level (%) 45.07 45.06 45.05 45.06 45.06
Calculated Lipid Level (%) 12.44 12.43 12.44 12.43 12.43