Black soldier fly meal in feed for Atlantic salmon - Erik-Jan Lock, Havforsknings Instituttet
1. Black soldier fly meal in feed for Atlantic salmon
- nutritional qualities and safety considerations -
Erik-Jan Lock, Irene Biancarosa, Ikram Belghit
2. Institute of Marine Research
• One of the largest Marine Research Institutes in Europe (~1000
employees)
• Main activities: research, advisory work and monitoring
3.
4. Institute of Marine Research
• One of the largest Marine Research Institutes in Europe (~1000
employees)
• Main activities: research, advisory work and monitoring
• Leading supplier of knowledge relating to the sustainable management of
the resources in our marine ecosystems and the whole food chain from the
sea to the table
8. Insect as feed ingredients
Black soldier fly (BSF) larvae
0
10
20
30
40
50
60
CP Arg His Ile Leu Lys Met Thr Val
Partially defatted BSF larvae
Atlantic salmon requirements (NRC, 2011)
Most of the indispensable amino acids are close to the requirements in Atlantic salmon
Amino acids composition= % of crude protein
Atlantic salmon requirements
10. Insect as natural feed ingredients for sustainable salmon
?
Evaluation of seaweed as a feed ingredients
for BSF larvae
Belghit I, Liland NS, Waagbø R, Biancarosa I, Pelusio N, Li Y, Krogdahl Å, Lock EJ. 2018. Potential of insect-based diets for
Atlantic salmon (Salmo salar). Aquaculture 491: 72-81
11. Increased EPA (retention around 35%)
Increased iodine
Increased tocopherols
N.S. Liland, I. Biancarosa, P. Araujo, D. Biemans, C.G. Bruckner, R. Waagbø, B.E. Torstensen, E.-J. Lock, Modulation of nutrient
composition of black soldier fly (Hermetia illucens) larvae by feeding seaweed-enriched media, PLOS ONE, 12 (2017)
I. Biancarosa, N.S. Liland, D. Biemans, P. Araujo, C.G. Bruckner, R. Waagbø, B.E. Torstensen, E.-J. Lock, H. Amlund, Uptake of heavy
metals and arsenic in black soldier fly (Hermetia illucens) larvae grown on seaweed-enriched media, J. Sci. Food Agric.
Black soldier fly larvae experimental trial
13. Insect as natural feed ingredients for sustainable salmon
?
Evaluation of insect meal (IM) as a feed
ingredients for Atlantic salmon
Belghit I, Liland NS, Waagbø R, Biancarosa I, Pelusio N, Li Y, Krogdahl Å, Lock EJ. 2018. Potential of insect-based diets for
Atlantic salmon (Salmo salar). Aquaculture 491: 72-81
14. Insect meal (IM)BSF larvae
FM/VP
Control diet (IM-0%)
66%IM
/VP
IM-66%
100%IM
/VP
IM-100%
Cargill
Replacement
60:40
Feed ingredients and experimental diets
Belghit et al
IM-33%
33%IM
/VP
Feeding period:16 weeks
WP2
Experiment 2. Grow-out trial
15. IM -0 IM -6 6IM -1 0 0 IM -0 IM -6 6IM -1 0 0 IM -0 IM -6 6IM -1 0 0
0
1 0 0 0
2 0 0 0
3 0 0 0
4 0 0 0
5 0 0 0
Grwoth(g)
Main results
• 3 fold increase in body weight
• No significant effects on growth
between groups
• No effects on nutrient digestibility
• Whole body FA composition was
affected by IM dietary inclusion
0 33 66 100 0 33 66 100 0 33 66 100
Belghit et al., submitted
WP2
Experiment 2. Grow-out trial
16. Head Kidney (HK) Isolated leukocytes
+ LPS
+ Poly I:C
+ Control (non treatment)
BSF insect meal (IM)
IM-0%
IM-66%
IM-100%
Gene expression
3rd October ≈2500 g
54 days
7th August ≈1200 g Isolated for 24h and added
treatment for additional 24h
2 fish/ cages = 6 fish/diet
IM -0 IM -6 6IM -1 0 0 IM -0 IM -6 6IM -1 0 0 IM -0 IM -6 6IM -1 0 0
0
1 0 0 0
2 0 0 0
3 0 0 0
4 0 0 0
5 0 0 0
Grwoth(g)
0 33 66 100 0 33 66 100 0 33 66 100
WP2
Experiment 2. Grow-out trial
Inflammatory related gene expression
Eicosanoid related gene expression
Stress related gene expression
17. Main results
WP2
Experiment 2. Grow-out trial
• Replacement of fishmeal with IM in the diets of Atlantic
salmon had no effect on the transcription of pro-
inflammatory genes in the head kidney cells.
• Fish fed with IM had down regulated transcription of
antioxidant and stress related genes.
IM - 0 IM - 66 IM - 100
0 .0
0 .2
0 .4
0 .6
0 .8
1 .0
S O D
¤
z
y
y
IM - 0 IM - 66 IM - 100
0 .0
0 .2
0 .4
0 .6
0 .8
1 .0
5 -LO X
z
y y
In vitro bacterial and viral response in head kidney leukocytes of Atlantic
salmon (Salmo salar) fed dietary insects meal
18. Sensory testing
• Softer texture of the raw fillets in fish
fed 100% IM than fish fed 33 % IM
• Higher rancidity odor of baked fillet
from fish fed 100% IM than the
control group
• No differences in color scores of raw
fish
Belghit et al., submitted
WP2
Experiment 2. Grow-out trial
19. Insect as feed ingredients for Atlantic salmon
Christopher Haatuft
20. -> Must comply with the associated legal constraints to
guarantee their safe use in fish feed ingredients
+ Large variety in origin and quality of insect products
Objective
Analytical approaches are urgently required for a proper detection
and identification of insect species-containing feed ingredients
ENTOFÔR project - From waste to resource
21. Insect meal (IM)
- 18 commercial IM samples
- Different companies - Different processing:
whole larvae meal or defatted insect meal
8 Black soldier fly (Hermetia illucens) (BSF)
5 Yellow mealworm (Tenebrio molitor) (YM)
3 Lesser mealworm (Alphitobius diaperinus) (LM)
2 House cricket (Acheta domesticus) (HC)
Materials
22. The protein content of the 18 IM samples
BSF YM LM HC
True protein content was determined as sum of amino acid residue
%oftotalAAAmino acid composition, protein content, and nitrogen-to-protein conversion factors of 21 seaweed species from Norwegian waters,
Biancarosa et al., 2016
0
2 0
4 0
6 0
8 0
1 0 0
0
2 0
4 0
6 0
8 0
1 0 0
Crude protein from overall nitrogen content, using the traditional
nitrogen-to-protein conversion factor of 6.25
BSF YM LM HC
Crudeprotein
23. Principal component analysis and hierchical clustering of the amino acids calculated
from 18 insect meal samples
BSF
HC
LM
YM
His
Glu
Met
Asp
Phe
Lys
Thr
Tau
Arg
Tyr
Iso
Ser
Pro
Ala
Val
Gly
Leu
BSF
YMLM
HC
BSF
25. PeptidesProteins extraction
Peptidomic profiling of different (commercially) available insect species
LC-MS/MS Spectral
Species Protein Peptides
BSF 1
21 53
BSF 2
43 196
BSF 3
21 103
BSF 4
81 275
BSF 5
18 91
BSF 6
58 303
BSF 7
56 156
BSF 8
55 220
YW 9
127 765
YW 10
138 689
YW 11
133 697
YW 12
54 235
YW 13
130 664
LW 14
94 509
LW 15
90 477
LW 16
94 389
HC 17
77 605
HC 18
75 454
Name Common names
Number of entries
(UNIPROT)
Number of proteins
(NCBI)
ACHETA DOMESTICUS House cricket 159 168
HERMETIA ILLUCENS Black soldier fly, BSF 56 348
TENEBRIO MOLITOR Yellow mealworm, beetles 527 607
ALPHITOBIUS
DIAPERINUS
Lesser mealworms,
buffalo worms, litter
beetles
27 63
ZOPHOBAS MORIO or
ZOPHOBAS ATRATUS
or ZOPHOBAS
RUGIPES
Morio worms, super
worm, king worm
19 38
26. PeptidesProteins extraction
Peptidomic profiling of different (commercially) available insect species
LC-MS/MS Spectral
All tandem mass spectra ≈ 4000
Pairwise comparaison CompareMS2
Records the similarity of the best matching spectrum in the corresponding range
Creating a distance matrix
Phylogenetic tree of the samples
27. Hierarchical Clustering of insect meal Proteomes
LM
LM
LM
YM
YM
YM
YM
YM
HC
HC
BSF
BSF
BSF
BSF
BSF
BSF
BSF
BSF
The phylogenetic analysis distinguished and successfully arranged the 18 different IM samples
species in groups of closely related families based on the comparison of 4000 tandem mass spectra+ increase -> spectral library data base
30. Developing peptide markers for the quantitative species specific
differentiation of insect meal
By using the gel-free shotgun proteomics approach in combination
with direct spectral comparison we were able to differentiate the
insect meal samples according to the taxonomic classification of the
insect species
Conclusion
By including IM in the diets, the concentration of lauric acid (12:0) increased (0.5-1.6% of the whole body total FA in the IM fed fish), while this FA was below the quantification limit in the whole fish fed diets without IM (IM0) (Table 6). The concentration of EPA and DHA as well as the n-3/n-6 ratio increased significantly with increasing inclusion of IM at 66% (IM66) and 100% (IM100) in the diets compared to fish fed with dietary IM0 and IM33. (Table 6).
The aim of the current study was to test how a partial or total replacement of dietary fishmeal with insect meal affect gene responses involved in inflammation and signaling pathways in Atlantic salmon (Salmo salar) head kidney leukocytes after exposure to bacterial or viral mimics.
bacterial mimic lipopolysaccharide (LPS) or viral mimic polyinosinic acid: polycytidylic acid (poly I: C) to induce an inflammatory response.
Different FA composition in the diets; migth have have changed the composition of cell membrane?
Have the FA profile in IM based diet caused changes in leukotriene production?
Less oxidative stress in fish given IM based diet?
Spider web diagram of the sensory evaluation of (A) raw and (B) baked fillets from IM0, IM33, IM66 and IM100 fed Atlantic salmon. A 100% substitution of fishmeal with insect meal (IM100) resulted in a softer texture of the raw fillets compared with the fillets of fish fed with only 33% fishmeal replacement (IM33). The rancidity odor of baked fillet from the IM100 group was significantly higher than in the control group without insect meal (IM0). p < 0.05; (one-way ANOVA). IM0 = diet without insect meal (IM) inclusion; IM33, IM66 and IM100 = 33, 66 and 100% replacement level of FM with IM, respectively.
Increasing number of insect-rearing farm for commercial purposes both inside and outside Europe
At least 26 companies in the world Example: ALPARE in Colombia produce BSF larvae 6 tons of IM/day
To demonstrate the feasibility of such a method we attempted to reconstruct a phylogenetic tree for humans and our closest living relatives based solely on uninterpreted tandem mass spectrometry data.