Complete replacement of soybean meal with black soldier fly larvae meal in

feeding program for broiler chickens from placement through to 49 days of age
reduced growth performance and altered organs morphology

Hannah Facey ,1 Munene Kithama , Mohsen Mohammadigheisar ,2 Lee-Anne Huber,

Anna K. Shoveller ,1 and Elijah G. Kiarie 3

Department of Animal Biosciences, University of Guelph, Guelph, N1G 2W1 ON, Canada

ABSTRACT Black soldier fly larvae meal (BSFLM) is and 49. On d 10, birds fed diets 12.5, 25, and
characterized with good nutritional and functional 0 + AGP had higher BWG than birds fed diets 0, 50,
attributes. However, there is limited data on inclusion of and 100 (P < 0.01), and birds fed diet 100 had lower
BSFLM in broiler chicken rations from placement BWG than birds fed diets 0 or 50 (P < 0.01). Birds
through to market weight. Therefore, we examined fed diets 50 and 100 had lower BWG than birds fed
growth and organ responses of partial to complete all other diets on d 24 and 49 (P < 0.05). Overall (d 0
replacement of soybean meal (SBM) with BSFLM in a −49), BSFLM linearly (P < 0.01) decreased BW,
practical feeding program. A total of 1,152 d-old male BWG, and FI and increased FCR and mortality. The
Ross £ Ross 708 chicks were allocated to 48 pens and overall BWG of 50 and 100% BSFLM birds was 92
assigned one of six diets (n = 8). The diets were: a basal and 81% of birds fed 0%BSFLM, repectively and core-
corn-SBM diet (0%BSFLM), 4 diets in which SBM in sponding overall FI was 96 and 90%. An increase in
0%BSFLM was replaced with BSFLM at 12.5, 25, 50, gizzard, small intestine, pancreas, and liver relative
and 100% and a final diet (0 + AGP) in which weights were observed with increasing BSFLM inclu-
0%BSFLM was treated with coccidiostat (70 mg Nara- sion (P < 0.01). The data indicated that lower levels
sin/kg) and antibiotic (55 mg Bacitracin Methylene Dis- of BSFLM could provide some growth-promoting
alicylate/kg). For energy fortification, soy oil was used effects commensurate to antibiotics in the starter
for 0%BSFLM diets and black soldier fly oil in the other phase. However, replacing SBM with greater amounts
diets. Body weight, feed intake (FI), BW gain (BWG), (≥50) of BSFLM reduced growth and increased organ
and mortality-corrected feed conversion ratio (FCR) size.
were reported. Organ weights were recorded on d 24
Key words: broiler chickens, black soldier fly larvae meal, growth performance, soybean meal replacement
2023 Poultry Science 102:102293
https://doi.org/10.1016/j.psj.2022.102293

INTRODUCTION Makkar et al., 2014). The BSFLM is a dense source of

Black soldier fly larvae meal (BSFLM) has been protein, fat, metabolizable energy (AME), phosphorus,
identified as a possible source of protein and energy in and fiber (Mwaniki and Kiarie, 2018). As part of the
broiler chicken diet which can be a replacement for com- fiber and fat components, BSFLM also contains high
mon plant-based proteins, such as soybean meal (SBM; concentrations of chitin and medium-chain fatty acids
(MCFA), such as lauric and myristic acid, which are
thought to improve both gut and immune health in
Ó 2022 The Authors. Published by Elsevier Inc. on behalf of Poultry
Science Association Inc. This is an open access article under the CC
broiler chickens through prebiotic and antimicrobial
BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/ properties (D€orper et al., 2020). These properties could
4.0/). reduce the reliance on antibiotics and coccidiostats in
Received June 9, 2022. the poultry industry (Bean-Hodgins and Kiarie, 2021).
Accepted October 22, 2022.
Presented in part at the Virtual 2021 Poultry Science Association In addition to the nutritional and functional benefits of
Annual Meeting, July 19-22, 2021. BSFLM, its use in broiler feed could help reduce the
1
Present address: Gowans Feed Consulting, New Hamburg, ON, environmental footprint of poultry feed production and
Canada, N3A 3H2. close the gap in a circular food economy (Barrera and
2
Present address: New-Life Mills, A division of Parrish & Heim-
becker, Cambridge, ON, Canada, N1T 2H9. Hertel, 2021) as black soldier fly larvae can consume
3
Corresponding author: ekiarie@uoguelph.ca

1
2 FACEY ET AL.

food waste and convert it into high-quality protein and 21°C by d 30 (Aviagen, 2014). Birds were exposed to
energy (Ravi et al., 2020). fluorescent lighting with 23 h of light (20+ lux) for the
However, within the current literature, growth perfor- first 4 d and then 16 h light: 8 dark (10−15 lux) light
mance responses have been variable for the inclusion of cycle for the remainder of the experiment in accord with
BSFLM in broiler diets. Previous researchers that Arkell Poultry Research Station standard operating pro-
included BSFLM below »30% replacement of SBM cedures.
observed no change or improved growth performance of
broilers, including BW, feed intake (FI), and feed con-
version ratio (FCR) (Cullere et al., 2016; Dabbou et al., Experimental Diets
2018; Onsongo et al., 2018; Schiavone et al., 2019; de
Partially defatted BSFLM (12.7% crude fat and
Souza Vilela et al., 2021). However, some studies imple-
52.5% CP, as-fed) and black soldier fly larvae oil
menting BSFLM levels greater than »30% replacement
(BSFLO) were obtained from a commercial manufac-
of SBM, more notably with levels above »50%, resulted
turer and vendor (Enterra feed Corp., Vancouver, BC,
in reduced growth performance through lower BW, FI,
Canada). The meal was a dry, ground product derived
and FCR as compared to lower inclusion rates (Dabbou
from black soldier fly (Hermetia illucens) larvae reared
et al., 2018; Onsongo et al., 2018; Velten et al., 2018;
on preconsumer recycled food collected from local farms,
Schiavone et al., 2019; Murawska et al., 2021). Addition-
food processors, and grocery stores. Coefficients for stan-
ally, results on organ weights and organ histology when
dardized ileal digestible (SID) amino acids and AME
BSFLM is included in higher amounts (>50% inclusion)
values for BSFLM were obtained from published litera-
are also variable. Some researchers did not observe any
ture (de Marco et al., 2015; Schiavone et al., 2017; Mwa-
changes in organ weights or organ tissue histology
niki and Kiarie, 2018) and the other feedstuffs from
(Onsongo et al., 2018; Velten et al., 2018; Schiavone
Evonik Aminodat 5.0 for diet formulation (Evonik
et al., 2019), whereas Murawska et al. (2021) observed
Nutrition and Care GmbH., 2016). The AME value of
an increase in liver weight when replacing 75% SBM
BSFLO was assumed to be equivalent to that of soybean
with BSFLM. Changes to organ weight are indicative of
oil (SBO) as research has shown no effect on growth
further health concerns, and therefore can aid in deduc-
performance when SBO is completely replaced with
ing information on the impact of BSFLM on broiler
BSFLO (Kim et al., 2020; Heuel et al., 2021). Diets were
chickens.
formulated for a 3-phase feeding program: Starter; d 0
The hypothesis of the study was that broilers will
to 10, Grower: d 11 to 24, and Finisher; d 25 to 49
experience a quadratic effect on growth performance
(Tables 1−3) and met or exceeded specifications for
where (1) feeding BSFLM will have commensurate
Ross x Ross 708 (Aviagen, 2014). In each phase, a
growth responses to antibiotic growth promoting
0%BSFLM corn-SBM-SBO basal diet was formulated
(AGP) diet via the functional properties of BSFLM and
with no prebiotic, probiotic, anticoccidial, or antimicro-
where (2) high levels of BSFLM will depress growth per-
bial growth-promoting substances added. Four BSFLM
formance due to decreased feed intake. The objective
diets were formulated based on the basal diet by replac-
was to examine the influence of partial to complete
ing SBM with increasing proportions of BSFLM: 12.5,
replacement of SBM with BSFLM in a broiler chicken
25, 50, and 100%. The 0%BSFLM diet was fed without
feeding program from placement through d 49 of age on
or with (0 + AGP) coccidiostat (Monteban 100, 70 mg
growth performance and organ weights.
Narasin/kg, Elanco Canada Ltd, Guelph, ON) and anti-
biotic growth promoter (BMD 110 G, 55 mg Bacitracin
Methylene Disalicylate/kg, Zoetis Canada Inc., Kirk-
MATERIALS AND METHODS land, QC). The 0%BSFLM diets used SBO and the
Animal care and use protocols are approved by the BSFLM diets used BSFLO for dietary energy fortifica-
University of Guelph Animal Care and Use Committee tion. The diets were mixed separately, with flushing
(AUP#4403) and birds were cared for in accordance between diets and were prepared in mash form. Repre-
with the Canadian Council on Animal Care guidelines sentative samples of the experimental diets, BSFLM,
(Canadian Council on Animal Care, 2009). SBM, BSFLO, and SBO were collected and immediately
shipped for chemical analyses and another portion was
stored at
20°C for future analyses.
Birds and Housing
A total of 1,152 d old (male) Ross x Ross 708 broiler Experimental Procedures
chicks were obtained from a commercial hatchery
(Maple Leaf Foods, New Hamburg, ON, Canada). The The 6 diets were allocated to 48-floor pens in 4 rooms
chicks were allocated to 48-floor pens (24 birds per pen) (12 pens per room) in a completely randomized block
according to hatch weight. The pens were in 4 separate (room) design (n = 8) and fed for 49 d. Birds had free
environmentally controlled rooms with 12 pens each, access to water and feed throughout the experiment.
providing 4.27 m2 of area per pen and fresh wood shav- The BW and FI were recorded at the end of each phase
ings. The room temperature was set to the breeder rec- (d 0, 10, 24, and 49) on a pen basis, and mortalities were
ommendation of 32°C on d 0 and gradually decreased to recorded throughout the trial for calculation of
 BLACK SOLIDER FLY MEAL FOR BROILER CHICKENS 3
Table 1. Composition of starter diets, as fed basis.

Starter phase, d 0−10

SBM replacement with BSFLM, %
Item 0 + AGP1 0 12.5 25 50 100
Corn 33.7 33.8 35.8 37.8 42.0 46.5
Wheat 20.0 20.0 20.0 20.0 20.0 20.0
Soybean meal 32.6 32.6 27.4 22.2 11.6 -
Pork meal 4.00 4.00 4.00 4.00 4.00 4.00
BSFLM2 - - 4.10 8.10 16.3 25.3
BSFLO2 - - 4.66 3.80 1.99 -
Soy oil 5.54 5.54 - - - -
L-Lysine HCL 0.35 0.35 0.41 0.48 0.61 0.76
DL-Methionine 0.38 0.38 0.40 0.43 0.48 0.53
L-Threonine 0.18 0.18 0.20 0.22 0.26 0.30
L-Tryptophan - - - 0.01 0.07 0.13
Limestone 0.68 0.68 0.6 0.52 0.36 0.18
Monocalcium phosphate 1.17 1.17 1.16 1.14 1.10 1.06
Salt 0.23 0.23 0.21 0.18 0.14 0.09
Sodium bicarbonate 0.09 0.09 0.10 0.10 0.12 0.13
Vitamin and trace minerals premix3 1.00 1.00 1.00 1.00 1.00 1.00
Antibiotic 0.05 - - - - -
Coccidiostat 0.07 - - - - -
Total 100 100 100 100 100 100
Calculated provisions
AMEn, kcal/kg 3,086 3,086 3,086 3,086 3,086 3,086
Crude fat, % 7.70 7.70 7.40 7.20 6.70 6.10
Linoleic acid, % 3.90 3.90 3.50 3.10 2.20 1.20
Crude protein, % 23.0 23.0 23.0 23.0 23.0 23.0
SID4 Lys, % 1.28 1.28 1.28 1.28 1.28 1.28
SID Met, % 0.67 0.67 0.69 0.70 0.73 0.77
SID Met + Cys, % 0.95 0.95 0.95 0.95 0.95 0.95
Calcium, % 0.96 0.96 0.96 0.96 0.96 0.96
Available P, % 0.48 0.48 0.48 0.48 0.48 0.48
Sodium, % 0.16 0.16 0.16 0.16 0.16 0.16
1
Coccidiostat (Monteban 100, 70 mg Narasin/kg, Elanco Canada Ltd, Guelph, ON) and antibiotic (BMD 110 G, 55 mg Bacitracin Methylene Disalicy-
late/kg, Zoetis Canada Inc., Kirkland, QC).
2
BSFLM; black soldier fly larvae meal, BSFLO; black solider fly larvae oil.
3
Provided per kg of premix: vitamin A (retinol), 880 KIU; vitamin D3 (cholecalciferol), 330 KIU; vitamin E, 4,000 IU; vitamin K3 (menadione), 330 mg;
vitamin B1 (thiamin), 400 mg; vitamin B2 (riboflavin), 800 mg; vitamin B3 (niacin), 5,000 mg; vitamin B5 (pantothenic acid), 1,500 mg; vitamin B6 (pyri-
doxine), 300 mg; vitamin B9 (folic acid), 100 mg; vitamin B12 (cyanocobalamin), 1200 mcg; biotin, 200 mcg; choline, 60,000 mg; Fe, 6000 mg; Cu, 1000
mg; I, 1 mg, Se, 30 mg.
4
SID: standardized ileal digestible.

mortality corrected FCR. The BWG was calculated Van Soest et al. (1991). Chitin concentration was deter-
from recorded BW on a pen basis at the end of each mined in the BSFLM samples by subtracting the ADF-
phase. On d 24 and 49, two birds per pen were randomly linked protein from the ash-free ADF as described by
selected and sacrificed via cervical dislocation for giz- Marono et al. (2015). Fatty acid concentration in
zard, small intestine, pancreas, spleen, liver, and bursa BSFLO and SBO samples was determined in a commer-
weights. cial lab (Activation Laboratories, Ancaster, ON) accord-
ing to O’Fallon et al. (2007). The amino acid contents of
BSFLM, SBM, and diet samples were also analyzed by
Sample Processing and Laboratory acid hydrolysis and oxidative hydrolysis (AOAC Inter-
Analyses national, 2005; Methods 994.12), followed by ultra-per-
formance liquid chromatography (UPLC; Waters
The BSFLM, SBM, and diet samples were finely Corporation, Milford, MA).
ground (CBG5 Smart Grind, Applica Consumer Prod-
ucts Inc., Shelton, CT) and thoroughly mixed prior to
analyses. Samples were submitted to a commercial lab
(SGS Canada Inc, Guelph, ON, Canada) for dry matter,
crude protein, crude fat, starch, and minerals analyses
Calculations and Statistics
(AOAC International, 2005; Methods 930.15, 935.11, Mortality corrected FCR is an adjusted FCR value
920.39, 920.40, 935.12). Feed samples were analyzed for which accounts for dead birds removed from the pens
gross energy (GE) using a bomb calorimeter (IKA Calo- throughout the trial. Mortality corrected FCR was cal-
rimeter System C 6000; IKA Works, Wilmington, NC) culated using the following equations.
using benzoic acid as the calibration standard. Neutral
detergent fiber (NDF) concentrations were determined 1. Mortality BWG = [(deadBW1 − BW/bird/pen at
using ANKOM 200 Fibre Analyzer (ANKOM Technol- the start of phase) + (deadBW2 − BW/bird/pen at
ogy, Fairport, NY) using the methodology described by the start of phase) + . . .]
4 FACEY ET AL.

Table 2. Composition of grower diets, as fed basis.

Grower phase, d 11−24

SBM replacement with BSFLM, %
Item 0 + AGP1 0 12.5 25 50 100
Corn 40.0 40.2 41.8 43.5 47.0 50.6
Wheat 20.0 20.0 20.0 20.0 20.0 20.0
Soybean meal 27.2 27.2 22.8 18.5 9.70 -
Pork meal 5.00 5.00 5.00 5.00 5.00 5.00
BSFLM2 - - 3.40 6.8 13.6 21.1
BSFLO2 - - 3.70 2.97 1.47 -
Soy oil 4.43 4.43 - - - -
L-Lysine HCL 0.31 0.31 0.37 0.42 0.54 0.66
DL-Methionine 0.33 0.33 0.35 0.37 0.42 0.46
L-Threonine 0.16 0.16 0.17 0.18 0.22 0.25
L-Tryptophan - - - - 0.05 0.11
Limestone 0.42 0.42 0.35 0.29 0.15 -
Monocalcium phosphate 0.73 0.73 0.71 0.70 0.67 0.64
Salt 0.23 0.23 0.22 0.20 0.16 0.12
Sodium bicarbonate 0.06 0.06 0.07 0.07 0.09 0.10
Vitamin and trace minerals premix3 1.00 1.00 1.00 1.00 1.00 1.00
Antibiotic 0.05 - - - - -
Coccidiostat 0.07 - - - - -
Total 100 100 100 100 100 100
Calculated provisions
AMEn, kcal/kg 3,100 3,100 3,100 3,100 3,100 3,108
Crude fat, % 6.90 6.90 6.70 6.60 6.10 5.80
Linoleic acid, % 3.40 3.40 3.10 2.70 2.00 1.20
Crude protein, % 21.5 21.5 21.5 21.5 21.5 21.5
SID4 Lys, % 1.15 1.15 1.15 1.15 1.15 1.15
SID Met, % 0.61 0.61 0.63 0.64 0.66 0.69
SID Met + Cys, % 0.87 0.87 0.87 0.87 0.87 0.87
Calcium, % 0.87 0.87 0.87 0.87 0.87 0.87
Available P, % 0.44 0.44 0.44 0.44 0.44 0.44
Sodium, % 0.16 0.16 0.16 0.16 0.16 0.16
1
Coccidiostat (Monteban 100, 70 mg Narasin/kg, Elanco Canada Ltd, Guelph, ON) and antibiotic (BMD 110 G, 55 mg Bacitracin Methylene Disalicy-
late/kg, Zoetis Canada Inc., Kirkland, QC).
2
BSFLM; black soldier fly larvae meal, BSFLO; black solider fly larvae oil.
3
Provided per kg of premix: vitamin A (retinol), 880 KIU; vitamin D3 (cholecalciferol), 330 KIU; vitamin E, 4,000 IU; vitamin K3 (menadione), 330 mg; vitamin
B1 (thiamin), 400 mg; vitamin B2 (riboflavin), 800 mg; vitamin B3 (niacin), 5,000 mg; vitamin B5 (pantothenic acid), 1,500 mg; vitamin B6 (pyridoxine), 300 mg;
vitamin B9 (folic acid), 100 mg; vitamin B12 (cyanocobalamin), 1200 mcg; biotin, 200 mcg; choline, 60,000 mg; Fe, 6000 mg; Cu, 1000 mg; I, 1 mg, Se, 30 mg.
4
SID: standardized ileal digestible.

2. Survivors BWG = (Total pen BW at the end of the RESULTS

current phase) − (Total pen BW at the end of the
previous phase)
Analyzed Chemical Composition
3. Mortality corrected FCR = Total FI / (Survivors Black solider fly larvae meal had a greater concen-
BWG + Mortality BWG) tration of crude protein, crude fat, phosphorus, cal-
cium, magnesium, and sodium on a DM basis
All the data were analyzed with the following statisti- (Table 4). The BSFLM also had a greater concentra-
cal model using the SAS Studio OnDemand for Academ- tion of starch and NDF than SBM, whereas SBM
ics software (2022): had a greater concentration of potassium. Addition-
ally, of the indispensable amino acids, BSFLM had
Y ¼ m þ trmti þ blockj þ eij
greater concentrations of His, Val, and Met, whereas
where Y represents the outcome, m is the mean of the SBM was greater in Arg and Trp. Of the dispensable
given observations (fixed effect), trmti is the effect amino acids, BSFLM was greater in Ala, Gly, and
of the inclusion level of BSFLM (fixed effect), blockj is Pro, and SBM was greater in Asp, Cys, and Glu.
the effect of block design, hence the variation between The fatty acid profile of BSFLO and SBO are pre-
the four rooms (random effect), and eij accounts for the sented in Table 5. The BSFLO and SBO had some
experimental error (random effect). The Proc GLIM- key differences in fatty acid profile, where BSFLO
MIX procedure was used to obtain the LS means and had greater total saturated fats and medium chain
Tukey Kramer’s post-hoc test was applied. The Proc fatty acids (MCFA) such as lauric and myristic
IML procedure was used to obtain contrast coefficients acid, as well as palmitic and palmitoleic acid.
(SAS Institute Inc, 2022). Coefficients were used in con- Conversely, SBO contained higher polyunsaturated
trast statements applied to the 0%BSFLM and BSFLM fatty acids such as stearic, linoleic, and linolenic
diets to identify linear and quadratic relationships acid. Within phase, the concentration for GE, CP,
resulting from the BSFLM inclusion. A P-value of ≤0.05 and crude fat were comparable; however, DM,
was considered significant. starch, and NDF increased with the level of BSFLM,
 BLACK SOLIDER FLY MEAL FOR BROILER CHICKENS 5
Table 3. Composition of finisher diets, as fed basis.

Finisher phase, d25−49

SBM replacement with BSFLM, %
Item 0 + AGP1 0 12.5 25 50 100
Corn 45.8 45.9 47.2 48.4 50.9 53.1
Wheat 20.0 20.0 20.0 20.0 20.0 20.0
Soybean meal 20.6 20.6 17.3 14.1 7.40 -
Pork meal 6.00 6.00 6.00 6.00 6.00 6.00
BSFLM2 - - 2.60 5.10 10.3 16.1
BSFLO2 - - 4.30 3.86 2.77 1.75
Soy oil 4.95 4.95 - - - -
L-Lysine HCL 0.33 0.33 0.37 0.41 0.50 0.59
DL-Methionine 0.31 0.31 0.33 0.34 0.38 0.41
L-Threonine 0.16 0.16 0.17 0.18 0.20 0.23
L-Tryptophan - - - 0.04 0.08
Limestone 0.18 0.18 0.13 0.08 - -
Monocalcium phosphate 0.33 0.33 0.31 0.30 0.33 0.52
Salt 0.23 0.23 0.21 0.20 0.17 0.14
Sodium bicarbonate 0.06 0.06 0.06 0.07 0.07 0.08
Vitamin and trace minerals premix3 1.00 1.00 1.00 1.00 1.00 1.00
Antibiotic 0.05 - - - - -
Coccidiostat 0.07 - - - - -
Total 100 100 100 100 100 100
Calculated provisions
AMEn, kcal/kg 3,200 3,200 3,200 3,200 3,200 3,200
Crude fat, % 7.80 7.80 7.60 7.50 7.20 7.10
Linoleic acid, % 3.80 3.80 3.50 3.30 2.70 2.20
Crude protein, % 19.5 19.5 19.5 19.5 19.5 19.5
SID4 Lys, % 1.03 1.03 1.03 1.03 1.03 1.03
SID Met, % 0.57 0.57 0.58 0.59 0.61 0.63
SID Met + Cys, % 0.80 0.80 0.80 0.80 0.80 0.80
Calcium, % 0.79 0.79 0.79 0.79 0.81 0.89
Available P, % 0.40 0.40 0.40 0.40 0.41 0.45
Sodium, % 0.16 0.16 0.16 0.16 0.16 0.16
1
Coccidiostat (Monteban 100, 70 mg Narasin/kg, Elanco Canada Ltd, Guelph, ON) and antibiotic (BMD 110 G, 55 mg Bacitracin Methylene Disalicy-
late/kg, Zoetis Canada Inc., Kirkland, QC).
2
BSFLM; black soldier fly larvae meal, BSFLO; black solider fly larvae oil.
3
Provided per kg of premix: vitamin A (retinol), 880 KIU; vitamin D3 (cholecalciferol), 330 KIU; vitamin E, 4,000 IU; vitamin K3 (menadione), 330 mg; vitamin
B1 (thiamin), 400 mg; vitamin B2 (riboflavin), 800 mg; vitamin B3 (niacin), 5,000 mg; vitamin B5 (pantothenic acid), 1,500 mg; vitamin B6 (pyridoxine), 300 mg;
vitamin B9 (folic acid), 100 mg; vitamin B12 (cyanocobalamin), 1200 mcg; biotin, 200 mcg; choline, 60,000 mg; Fe, 6000 mg; Cu, 1000 mg; I, 1 mg, Se, 30 mg.
4
SID: standardized ileal digestible.

Table 4. Analyzed composition of black soldier larvae meal

(BSFLM) and soybean meal (SBM) used in the study, DM basis. Table 5. Analyzed composition of black soldier larvae oil
(BSFLO) and soybean oil (SBO) used in the study, as fed basis.
Item BSFLM SBM
Dry matter, % 95.42 88.22 Fatty acids, % of total fat BSFLO SBO
Crude protein, % 55.1 51.4
Crude fat, % 13.3 2.64 Decanoic 0.73 0.00
Starch, % 3.73 1.29 Undecanoic 0.01 0.00
Chitin, % 6.67 0.00 Lauric 35.32 0.00
Neutral detergent fiber, % 19.7 6.90 Myristic 7.32 0.06
Gross energy, kcal/kg 5,341 4,746 Myristoleic 0.24 0.00
Calcium, % 1.51 0.14 Pentadecanoic 0.07 0.01
Phosphorus, % 0.70 0.62 Palmitic 12.08 9.93
Potassium, % 1.36 2.41
Palmitoleic 3.01 0.08
Magnesium, % 0.36 0.31
Sodium, % 0.18 0.05 Heptadecanoic 0.10 0.10
Indispensable Amino Acids, % cis-10-Heptadecenoic 0.06 0.05
Arg 2.75 3.55 Stearic 2.37 3.86
His 1.73 1.38 Oleic 20.02 18.83
Ile 1.99 1.97 Linolelic 13.00 55.35
Leu 3.60 3.79 Arachidic 0.11 0.29
Lys 2.75 2.86 g-Linolenic 0.05 0.03
Met 0.80 0.61
Gadoleic 0.17 0.19
Phe 2.73 2.65
Thr 1.95 1.84 Linolenic 2.50 8.76
Trp 0.72 1.40 Heneicosanoic 0.00 0.06
Val 3.14 2.35 cis-11,14-Eicosadienoic 0.02 0.06
Dispensable Amino Acids, % Behenic 0.03 0.31
Ala 3.38 2.13 Tricosanoic 0.02 0.00
Asp 4.91 5.77 Methyl cis-5,8,11,14-eicosatetraenoic 0.00 0.04
Cys 0.50 0.77 Lignoceric 0.01 0.10
Glu 5.45 7.54
Others 2.76 1.90
Gly 2.30 1.75
Pro 3.54 2.76 Total saturated fatty acids % 58.00 15.00
Ser 2.36 2.61 Total monounsaturated fatty acids, % 23.00 19.00
Tyr 1.78 1.61 Total poly unsaturated fatty acids, % 16.00 64.00
6 FACEY ET AL.

Table 6. Analyzed chemical composition of starter diets, as fed basis.

Starter phase, d 0−10

SBM Replacement with BSFLM,2 %
Item 01 12.5 25 50 100
Dry matter, % 89.4 89.7 89.7 90.1 90.6
Gross Energy, kcal/kg 4167 4210 4260 4221 4252
Crude protein, % 22.4 23.2 21.4 23.5 23.4
Crude fat, % 6.7 7.0 7.3 5.7 6.2
Starch, % 37.8 26.6 37.9 38.6 40.7
NDF,3 % 10.0 11.8 12.6 15.2 14.6
Calcium, % 0.73 0.80 0.83 0.91 1.02
Phosphorus, % 0.73 0.71 0.78 0.70 0.79
Potassium, % 0.89 0.89 0.87 0.77 0.67
Magnesium, % 0.17 0.18 0.18 0.18 0.19
Sodium, % 0.15 0.20 0.17 0.18 0.16
Amino acids, %
Indispensable
Arg 1.38 1.39 1.28 1.32 1.1
His 0.57 0.61 0.54 0.65 0.62
Ile 1.10 0.86 0.79 1.11 0.92
Leu 1.97 1.76 1.72 2.07 1.8
Lys 1.29 1.39 1.57 1.65 1.59
Met 0.55 0.59 0.68 0.76 0.77
Phe 1.34 1.12 0.67 1.31 0.96
Thr 0.87 1.07 0.98 0.99 1.04
Trp 0.47 0.46 0.44 0.37 0.32
Val 0.95 1.04 1.02 1.14 1.14
Dispensable
Ala 1.01 1.10 1.17 1.34 1.34
Asp 2.11 2.17 2.18 2.11 1.84
Cys 0.44 0.46 0.42 0.41 0.38
Glu 3.17 1.32 3.20 3.08 2.57
Gly 0.80 0.88 0.80 1.01 0.98
Pro 1.32 1.43 1.36 1.61 1.57
Ser 1.06 1.07 1.07 1.11 0.98
Tyr 0.43 0.53 0.17 0.64 0.41
1
Same analyzed nutrient composition as diet 0%BSFLM + AGP (Coccidiostat (Monteban 100, 70 mg Narasin/kg, Elanco Canada Ltd, Guelph, ON)
and antibiotic (BMD 110 G, 55 mg Bacitracin Methylene Disalicylate/kg, Zoetis Canada Inc., Kirkland, QC)).
2
BSFLM; black soldier fly larvae meal.
3
NDF; neutral detergent fibre.

and potassium decreased (Tables 6−8). One notable lowest for birds fed diet 0 + AGP (P < 0.01) and great-
difference is that the CP concentration in diet 0 in est for birds fed diets 0 and 50 (P < 0.01), while all other
the finisher phase was lower than expected. Tables 6 treatments were intermediate.
to 8 also presented concentrations of the amino acid On d 24, BW decreased linearly and quadratically as
in the starter, grower, and finisher phases, respec- the level of BSFLM increased (P < 0.01), where birds
tively. fed diets 0, 12.5, and 25 had greater BW than birds fed
diets 50 and 100 (P < 0.01). However, birds fed diets
12.5 and 25 had intermediate BW compared to birds fed
Growth Performance the 0 + AGP diet and diet 0. The BWG during the
grower phase decreased linearly as birds fed diets 0,
Both the d 10 BW and starter BWG decreased line- 12.5, and 25 had greater BWG than birds fed diets 50
arly and quadratically as the inclusion level of BSFLM and 100 (P < 0.01); no differences were observed for
increased (P < 0.01) (Table 9). A quadratic decrease BWG for birds fed diets 0 + AGP, 0, 12.5, and 25. The
occurred where birds fed diets 12.5 and 25 had higher FI decreased linearly with increasing BSFLM level as
BW than diets 0, 50, and 100 (P < 0.01), and a linear birds fed diets 0 and 12.5 had greater FI than birds fed
decrease occurred where diet 100 resulted in lower BW diet 100 (P < 0.01), and birds fed diets 25 and 50 were
than diets 0 and 50 (P < 0.01). On d 10, birds fed diets intermediate to birds fed diets 0, 12.5, and 100. The
12.5 and 25 had BW not different from birds fed the FCR increased linearly and quadratically with increas-
0 + AGP diet, whereas birds fed diets 0, 50, and 100 had ing BSFLM inclusion level as birds fed diet 12.5 had the
lower BW (P < 0.01). The FI also decreased linearly and lowest FCR and birds fed diet 100 had the greatest FCR
quadratically as the level of BSFLM increased (P < (P < 0.01) with all other diets intermediate.
0.01), where birds fed diets 0, 12.5, 25, and 50 had On d 49, BW decreased linearly as the dietary inclu-
greater FI than birds fed diet 100, and birds fed diet 12.5 sion level of BSFLM increased, where birds fed diets 0,
had greater FI than birds fed diets 50 and 100 (P < 12.5, and 25 had greater BW than birds fed diets 50 and
0.01). Compared to birds fed the 0 + AGP diet, only 100 (P < 0.01). Birds fed diet 0 + AGP had the greatest
birds fed diet 100 had lower FI (P < 0.05). The FCR was BW versus all other treatment groups (P < 0.01). The
 BLACK SOLIDER FLY MEAL FOR BROILER CHICKENS 7
Table 7. Analyzed chemical composition of grower diets, as fed basis.
Grower, d 11−24
SBM replacement with BSFLM2, %
Item 01 12.5 25 50 100
Dry matter, % 89.1 89.2 89.2 89.8 89.9
Gross Energy, kcal/kg 4191 4132 4189 4210 4184
Crude protein, % 19.2 19.4 20.9 21.5 20.6
Crude fat, % 7.1 6.6 5.7 5.9 5.6
Starch, % 39.6 39.6 42.1 42.7 44.7
NDF,3 % 12.2 11.7 14.1 14.2 15.7
Calcium, % 0.60 0.65 0.65 0.70 0.69
Phosphorus, % 0.56 0.54 0.58 0.59 0.56
Potassium, % 0.86 0.80 0.75 0.70 0.58
Magnesium, % 0.16 0.17 0.16 0.19 0.19
Sodium, % 0.14 0.16 0.17 0.15 0.14
Amino acids, %
Indispensable
Arg 1.28 1.20 1.19 1.13 0.99
His 0.54 0.51 0.54 0.55 0.52
Ile 0.86 1.01 0.94 0.96 0.75
Leu 1.71 1.94 1.85 1.87 1.64
Lys 1.12 1.28 1.37 1.34 1.38
Met 0.58 0.57 0.62 0.70 0.73
Phe 1.15 1.23 1.14 1.19 0.83
Thr 0.84 0.89 0.97 0.90 0.90
Trp 0.45 0.43 0.44 0.38 0.36
Val 0.87 0.87 0.89 0.91 0.95
Dispensable
Ala 0.96 1.07 1.13 1.25 1.30
Asp 1.89 1.98 1.96 1.84 1.71
Cys 0.46 0.44 0.44 0.35 0.35
Glu 2.93 3.04 2.98 2.75 2.49
Gly 0.79 0.80 0.82 0.99 0.91
Pro 1.27 1.34 1.37 1.51 1.45
Ser 0.98 1.00 1.01 0.97 0.93
Tyr 0.43 0.47 0.48 0.41 0.35
1
Same analyzed nutrient composition as diet 0%BSFLM + AGP (Coccidiostat (Monteban 100, 70 mg Narasin/kg, Elanco Canada Ltd, Guelph, ON)
and antibiotic (BMD 110 G, 55 mg Bacitracin Methylene Disalicylate/kg, Zoetis Canada Inc., Kirkland, QC)).
2
BSFLM: black soldier fly larvae meal.
3
NDF: neutral detergent fibre.

Table 8. Analyzed chemical composition of finisher diets, as fed basis.

Finisher, d 25−49
SBM Replacement with BSFLM2, %
Item 01 12.5 25 50 100
Dry matter, % 88.9 88.6 88.5 88.8 89.2
Gross Energy, kcal/kg 4231 4238 4231 4226 4372
Crude protein, % 17.0 19.0 20.4 19.3 18.6
Crude fat, % 7.8 7.5 6.9 7.3 6.5
Starch, % 41.7 41.3 41.4 43.4 46.8
NDF,3 % 12.3 12.7 12.2 14.5 15.2
Calcium, % 0.66 0.60 0.49 0.68 0.87
Phosphorus, % 0.55 0.55 0.47 0.57 0.63
Potassium, % 0.72 0.72 0.67 0.61 0.51
Magnesium, % 0.16 0.17 0.15 0.16 0.16
Sodium, % 0.14 0.17 0.14 0.14 0.16
Amino acids, %
Indispensable
Arg 1.16 1.14 1.14 1.03 0.89
His 0.49 0.50 0.51 0.50 0.49
Ile 0.85 0.82 0.70 0.63 0.59
Leu 1.75 1.77 1.60 1.48 1.39
Lys 1.06 1.17 1.40 1.22 1.28
Met 0.54 0.53 0.59 0.61 0.63
Phe 1.17 1.14 0.92 0.73 0.78
Thr 0.88 0.84 0.83 0.80 0.82
Trp 0.53 0.46 0.39 0.39 0.34
Val 0.75 0.82 0.94 0.90 0.90
Dispensable
Ala 0.98 1.06 1.12 1.11 1.13
Asp 1.75 1.84 1.86 1.65 1.47
Cys 0.34 0.34 0.31 0.27 0.23
Glu 2.82 2.81 2.81 2.48 2.19
Gly 0.84 0.85 0.86 0.85 0.82
Pro 1.29 1.34 1.35 1.35 1.32
Ser 0.95 1.00 0.94 0.87 0.80
Tyr 0.40 0.39 0.45 0.25 0.41
1
Same analyzed nutrient composition as diet 0%BSFLM + AGP (Coccidiostat (Monteban 100, 70 mg Narasin/kg, Elanco Canada Ltd, Guelph, ON)
and antibiotic (BMD 110 G, 55 mg Bacitracin Methylene Disalicylate/kg, Zoetis Canada Inc., Kirkland, QC)).
2
BSFLM: black soldier fly larvae meal.
3
NDF: neutral detergent fibre.
8 FACEY ET AL.

Table 9. Effects of replacing soybean (SBM) with increasing proportions of black soldier fly larvae meal (BSFLM) on growth perfor-
mance of broiler chickens.

SBM replacement with BSFLM,3 % Responses to BSFLM2

Item 0+AGP1 0 12.5 25 50 100 SEM P-value Linear Quadratic
Placement BW, g/bird 45.5 45.6 45.6 45.6 45.5 45.7 0.11 0.420 - -
Starter, d 0−10
BW, g/bird 265.4a 245.6b 259.9a 261.3a 244.2b 203.0c 2.99 <0.001 <0.001 <0.001
BW gain, g/bird 220.0a 200.2b 214.5a 215.9a 198.9b 157.4c 2.91 <0.001 <0.001 <0.001
Feed intake, g/bird 236.7ab 239.2ab 244.7a 235.0ab 230.0b 117.3c 4.11 <0.001 <0.001 <0.001
FCR 1.082b 1.197a 1.146ab 1.092b 1.162a 1.132ab 0.02 <0.001 0.149 0.081
Grower, d 11−24
Final BW, g/bird 973.3a 904.7b 926.6ab 921.8ab 818.0c 669.2d 19.82 <0.001 <0.001 0.008
BW gain, g/bird 709.6a 660.8a 668.3a 662.2a 575.5b 467.9c 17.09 <0.001 <0.001 0.116
Feed intake, g/bird 1,218a 1,208a 1,128a 1,124ab 1,106ab 1,009b 45.18 <0.001 <0.001 0.685
FCR 1.651bc 1.877ab 1.462c 1.714abc 1.741abc 1.963a 0.11 <0.001 0.008 0.031
Finisher, d 25−49
Final BW, g/bird 3,257a 2,996b 3,041b 3,067b 2,748c 2,449d 41.85 <0.001 <0.001 0.056
BW gain, g/bird 2,275a 2,082b 2,105b 2,137ab 1,921c 1,771c 37.96 <0.001 <0.001 0.317
Feed intake, g/bird 3,801a 3,625abc 3,680ab 3,720ab 3,520bc 3,377c 59.65 <0.001 <0.001 0.302
FCR 1.671b 1.743b 1.746b 1.741b 1.831a 1.909a 0.02 <0.001 <0.001 0.600
Overall, d 0−49
BW gain, g/bird 3,211a 2,950b 2,995b 3,022b 2,703c 2,403d 41.84 <0.001 <0.001 0.055
Feed intake, g/bird 5,258a 5,076ab 5,056ab 5,082ab 4,859bc 4,566c 110.6 <0.001 <0.001 0.336
FCR 1.668d 1.746c 1.711cd 1.714cd 1.815b 1.910a 0.02 <0.001 <0.001 0.042
Mortality, % 3.6b 1.2b 6.0b 1.8b 3.6b 18.5a 1.96 <0.001 <0.001 0.009
Within a row LSMeans with different superscripts differs, P < 0.05.
a-d
1
Coccidiostat (Monteban 100, 70 mg Narasin/kg, Elanco Canada Ltd, Guelph, ON) and antibiotic (BMD 110 G, 55 mg Bacitracin Methylene Disalicy-
late/kg, Zoetis Canada Inc., Kirkland, QC).
2
Contrasts in LSMeans tested among diets 0, 12.5, 25, 50, and 100
3
BSFLM: black soldier fly meal.

BWG during the finisher phase also decreased linearly as Overall (d 0−49), the BWG and FI decreased linearly
birds fed diets 0 and 12.5 had greater BWG than birds as birds fed diets 0, 12.5, and 25 had greater BWG than
fed diets 50 and 100 (P < 0.01), while birds fed diet 25 birds fed diets 50 and 100 (P < 0.01). Birds fed all other
were intermediate. Likewise, BWG for birds fed diet diet treatments had lower BWG than birds fed diet
0 + AGP was the greatest compared to all other diet 0 + AGP (P < 0.01). The overall FCR decreased linearly
treatments (P < 0.01). The FI decreased linearly where and quadratically where birds fed diets 0, 12.5, and 50
birds fed diets 12.5 and 25 had greater FI than birds fed had lower FCR than birds fed diets 50 and 100 (P <
diet 100 (P < 0.01). Birds fed diet 0 + AGP had greater 0.01), with birds fed 12.5 and 25 intermediate to birds fed
BW than birds fed diets 50 and 100 (P < 0.01). The diets 0 and 0 + AGP, given that birds fed diet 0 + AGP
FCR increased linearly where birds fed diets 0, 12.5, and had the lowest result. Overall, mortality increased line-
25 had lower FCR than birds fed diets 50 and 100 (P < arly and quadratically where birds fed diet 100 had
0.01). There was no difference between the 0+AGP diet greater mortality compared to all other diets and birds
and diets 0, 12.5, and 25. fed diets 0 and 25 had the lowest mortalities (P < 0.01).

Table 10. Effects of replacing soybean meal (SBM) with increasing proportions of black soldier fly larvae meal (BSFLM) on organ
weights (g/kg BW) and breast muscle attributes in broiler chickens.

SBM replacement with BSFLM, % Responses to BSFLM3

1
Item 0 + AGP 0 12.5 25 50 100 SEM P-value Linear Quadratic
24 d old
Gizzard 25.1ab 28.8a 23.7b 23.3b 25.4ab 26.8ab 0.90 0.001 0.644 0.002
Small intestine 29.6b 31.9ab 33.2a 31.5ab 31.2ab 34.5a 0.84 0.005 0.063 0.053
Pancreas 3.0c 3.2bc 3.2bc 3.3bc 3.5ab 3.8a 0.12 <0.001 <0.001 0.902
Liver 18.5c 21.1bc 22.8ab 20.8bc 21.1bc 25.4a 1.59 0.001 0.005 0.058
Spleen 0.8 0.9 0.9 0.8 0.8 0.8 0.05 0.879 0.320 0.505
Bursa 2.6 2.7 2.5 2.6 2.8 2.9 0.19 0.676 0.163 0.605
49 d old
Gizzard 16.8b 19.7ab 20.1ab 17.9ab 20.6a 19.7ab 0.81 0.014 0.696 0.998
Small intestine 16.7b 19.9a 18.9ab 19.2ab 19.5a 20.8a 0.90 0.002 0.110 0.193
Pancreas 2.0b 2.2ab 2.1ab 2.2ab 2.2ab 2.4a 0.13 0.015 0.032 0.405
Liver 20.5b 22.7ab 22.1b 23.0ab 24.1ab 26.2a 1.31 0.001 0.001 0.709
Spleen 1.1 1.0 1.1 1.1 1.0 1.2 0.07 0.502 0.101 0.392
Bursa 1.7 1.4 1.7 1.5 1.7 1.9 0.14 0.230 0.043 0.921
Within a row LSMeans with different superscripts differs, P < 0.05.
a-c
1
Coccidiostat (Monteban 100, 70 mg Narasin/kg, Elanco Canada Ltd, Guelph, ON) and antibiotic (BMD 110 G, 55 mg Bacitracin Methylene Disalicy-
late/kg, Zoetis Canada Inc., Kirkland, QC).
2
Contrasts in LS Means tested among diets 0, 12.5, 25, 50, and 100.
 BLACK SOLIDER FLY MEAL FOR BROILER CHICKENS 9
Relative Organ Weights microbiome (Metzer Farms, 2018) and must be eutha-
nized to avoid welfare issues when the feeders and water-
At the end of the grower phase (d 24), gizzard weight ers are adjusted according to bird growth.
quadratically increased among the BSFLM diets, where A study conducted by Dabbou et al. (2018) reported
birds fed diets 12.5 and 25 had the lowest gizzard quadratic responses in BW to increasing BSFLM level
weights (P < 0.01) (Table 10). Small intestine weight throughout the starter, grower, and finisher phases,
increased where birds fed the 0 + AGP diet had the low- with the highest BW observed for the 10% inclusion
est weight, birds fed the 12.5 and 100 diets had the high- diet, similarly to the quadratic decrease observed in the
est weights, and all other diets were intermediate starter phase in the present study. However, Dabbou
(P < 0.01). The pancreas weight increased linearly with et al. (2018) reported most statistically significant
increasing dietary BSFLM inclusion, where birds fed responses in the later phases (grower and finisher) as
diet 100 had the greatest pancreas weight, and birds fed opposed to the starter phase in the present study. Dab-
diets 0, 12.5, 25, and 50 were intermediate to birds fed bou et al. (2018) saw otherwise no change in BW, BWG,
diet 0 + AGP and diet 100 (P < 0.01). Birds fed diet FI, or FCR for diets including 5% and 10% BSFLM and
0 + AGP had a lower pancreas weight than birds fed reduced BW and FCR in the diet containing 15% inclu-
diets 50 and 100. The liver weight increased linearly sion of BSFLM compared to the control diet (0%
with increasing dietary BSFLM inclusion where birds BSFLM). It is thought that BSFLM and oil possess pre-
fed diets 0, 25, and 50 had lower liver weights than birds biotic and antimicrobial properties from the chitin and
fed diet 100, yet birds fed diet 12.5 were intermediate (P medium-chain fatty acid contents (D€orper et al., 2020);
< 0.01). Birds fed diet 0 + AGP had the lowest liver however, previous studies used partially defatted
weight (P < 0.01). The spleen and bursa were not influ- BSFLM without additional BSFLO (Dabbou et al.,
enced by dietary treatment. 2018), indicating that even relatively low amounts of
At the end of the finisher phase (d 49), birds fed diet chitin and MCFA can positively impact growth perfor-
0 + AGP had lower gizzard weight than birds fed diet mance to achieve BW not different from birds fed AGP
50, while all other diets were intermediate (P < 0.05), diets. Furthermore, Dabbou et al. (2018) reported an
and lower small intestine weight than birds fed diets 0, increase in FI only during the starter phase, compared
50, and 100, with intermediate values observed for diets to the present study, where we observed a decrease in FI
12.5 and 25 (P < 0.01). Pancreas weight increased line- with increasing BSFLM inclusion through all 3 phases.
arly among the BSFLM diets, where birds fed diet 100 The FI depression in the present study may help explain
had the highest pancreas weight and all other diets were the unsustained benefits of BSFLM in later phases,
intermediate to birds fed diets 0 + AGP and 100 (P < where Dabbou et al. (2018) observed maintained FI and
0.05). Liver weight also increased linearly among the thus sustained levels of functional compounds such as
BSFLM diets, where birds fed diet 12.5 had a lower liver chitin and MCFA. Likewise, de Souza Vilela et al.
weight than birds fed diet 100, while all other diets were (2021) fed broilers up to 20% inclusion of full-fat
intermediate (P < 0.01). Birds fed diet 0 + AGP only BSFLM and observed no change in growth performance
had a lower liver weight than birds fed diet 100 (P < in the starter phase, and a linear increase in BW, BWG,
0.01). Spleen weight did not change in response to and FCR in the grower and finisher phases with no
BSFLM inclusion; however, bursa weight increased line- change in FI. Additionally, Cullere et al. (2016) and
arly with inclusion of BSFLM (P < 0.05). Onsongo et al. (2018) reported no change in growth per-
formance parameters, including FI, when feeding
DISCUSSION broilers below 55.5% replacement of SBM with BSFLM
or BSF pupae meal. However, any additional benefits
The objective of the study was to examine the influ- were not reported despite these studies utilizing diets
ence of partial to complete replacement of SBM and with similar amounts of crude fat compared to the pres-
BSFLM in a broiler feeding program on growth perfor- ent study. The crude fat content is considered the source
mance. The diets were formulated based on book value of these benefits, given the fatty acid profile (MCFA).
specifications for ingredients. Key indispensable amino Cullere et al. (2016) and Onsongo et al. (2018) used
acids (Lys, Met, Met + Cys) were formulated to meet defatted BSF products, and supplemented with either
the requirements, and most other amino acid concentra- SBO or corn oil, rather than BSFLO and therefore
tions met or exceeded specifications. However, diet anal- would not have the functional benefits from high MCFA
yses revealed several amino acids that were rather low. as BSFLO does. Diets 0, and 25 had the lowest mortal-
For example, total Arg displayed consistent deficiencies ities. Additionally, several relative organ weights
among the diets relative to digestible Arg specifications increased with increasing BSFLM inclusion levels
of 1.40, 1.27, and 1.17% for starter, grower, and finisher, including the gizzard, small intestine, and liver. Given
respectively (Aviagen, 2014). Additionally, Thr, Val, this evidence, further research should be conducted on
and Ile also displayed deficiencies in some diets. The the use of BSFLM and oil as a supplement (<»25%) to
high mortalities in diet 100 was due to the increased replace antibiotics and coccidiostats in AGP diets.
need for euthanasia, due to mainly lameness in the form Current literature indicates that the overall linear
of leg problems, and non-uniform growth leading to decrease in performance observed throughout the study
more runts. Runts may be the result of a poor gut with high inclusion levels of BSFLM could be due to
10 FACEY ET AL.

greater fiber contents or limited available amino acid is possible that BSFLM can trigger increased bacterial
supply. Murawska et al. (2021) observed a negative lin- populations in the gut, causing the gut wall to thicken
ear relationship in BW, BWG, FI, and FCR in broilers via increased mucosa or muscularis production, resulting
fed 50%, 75%, and 100% replacement of SBM with in a greater overall weight.
BSFLM, and suggested the decreased performance Reduced nutrient absorption and digestibility would
mainly results from the reduction in FI. It is suggested also limit the amino acids available to the bird. A study
that reduced FI could be caused by impaired protein uti- conducted by Velten et al. (2018) replaced 50% SBM
lization linked to increased dietary chitin content when with BSFLM in broiler diets containing standard amino
BSFLM is included at higher levels in the diet. Chitin is acid supplementation and fortified amino acid supple-
representative of the fiber in BSFLM, and as seen in mentation (addition of Met, Lys, Thr, Arg, Val).
Murawska et al. (2021) and the present study, the NDF Reduced growth performance was observed when the
content of the diets increases as the level of BSFLM broilers received the standard amino acid supplementa-
inclusion increases. High fiber content or NDF, has been tion, but no change in performance parameters was
seen to increase anterior digestive tract weight (gizzard), observed when receiving supplemental amino acids, indi-
reduce digesta transit time, and subsequently reduce cating that the birds were not receiving sufficient amino
nutrient digestibility (Kras et al., 2013; Sanchez et al., acids in a standard diet with BSFLM inclusion. Simi-
2021). larly, it was recently demonstrated that the addition of
Therefore, given the effects seen on not only reduced indispensable synthetic amino acids in BSFLM diet to
FI, but also increased FCR in the present study, it is equal levels of amino acids in soybean meal diet
thought that the decreased BW and BWG is not only improved protein utilization efficiency in BSFLM
due to reduced FI, but also a reduction in nutrient (Cheng et al., 2022). In the present study, the diets were
digestibility resulting from high crude fiber, and could analyzed, and the most notable effect on amino acid con-
have further adverse effects on other areas of the body centrations was the Arg deficiency. This deficiency
such as feather development and organ function. increased as the level of BSFLM increased, correspond-
According to the current literature, the gizzard, primar- ing with lower levels of arginine in BSFLM versus SBM
ily functioning as the site for grinding and breaking diet. Furthermore, the Arg: Lys ratio was below the
down feed particles, is expected to increase in size and NRC recommendation of 0.9 to 1.18 (Balnave and
mass as larger particles and more structural and fibrous Barke, 2002) in diets 25, 50, and 100 for the starter and
material are added to the diet (Kiarie and Mills, 2019). finisher phases, and diet 100 for the grower phase. Argi-
An increase in gizzard weight was seen in diets contain- nine plays an essential role in broiler growth, feather
ing high levels of BSFLM (50 and 100), which also con- development, and immunity, and can influence the effec-
tained the highest concentration of chitin, and tiveness of all other amino acids in protein synthesis;
consequently NDF, suggesting that the high fiber con- however, is heavily influenced by Lys concentrations
tent in diets 50 and 100 caused the increase in gizzard (Balnave and Barke, 2002). The literature reviewed by
weight. In addition to reduced nutrient digestibility via Balnave and Barke (2002) suggest that Lys and Arg do
increased NDF, reduced nutrient absorption in the small not compete for absorption in the small intestine, but
intestine could also have contributed to poor perfor- instead compete for renal reabsorption. If the Arg: Lys
mance. In the present study, the small intestinal weight ratio is imbalanced towards Lys, excess Lys will be reab-
increased with increasing levels of BSFLM inclusion; sorbed, a process which is also influenced by potassium
however, on d 24 and 49, diet 0 + AGP had significantly levels, and will result in the overproduction of arginase
lighter small intestine compared to all other diets. Ear- by the kidney. Arginase activity is closely correlated to
lier research has shown that AGP inclusion causes intes- reduced FI and BWG in broilers as it also breaks down
tinal wall thinning (Coates et al., 1955). Thinned Arg, causing more of a deficiency (Balnave and Barke,
intestinal walls allow nutrients to pass more easily, 2002). This indicates that the reduced performance seen
increasing overall nutrient absorption and hence feed in Velten et al. (2018) could be caused by amino acid
utilization (Madge, 1969). The thinned intestinal wall imbalances resulting from the Arg: Lys ratio and could
likely led to the decrease in intestinal weight in the 0 be rectified with amino acid supplementation, or as also
+AGP fed birds. On the other hand, research has shown suggested by Balnave and Barke (2002), and potassium
that the protein source in poultry diets, notably vegeta- supplementation. Given that an Arg deficiency can
ble versus animal sources, can influence the microbiome reduce the effectiveness of other amino acids for use for
in the gut through increased microbial diversity (Bean- protein synthesis, a build up of unavailable amino acids
Hodgins and Kiarie, 2021). Animal proteins contain would require an increase of catabolism of amino acids
high amounts of crude protein and can cause the fermen- via the liver. Therefore, the liver would be working
tation of excess crude protein in the intestinal tract, harder to break down the excess amino acids (He et al.,
which feeds harmful bacteria such as C. perfringens 2021). The increased metabolic activity taking place in
(Drew et al., 2004; Wilkie et al., 2005; Bean-Hodgins the liver would cause the increase in size and relative
and Kiarie, 2021). Indeed, much of the protein in insect weight as seen in the present study. An alternative possi-
meal has been demonstrated to be bound in chitin and bility for hepatic hypertrophy is if amino acids are
concentration of chitin had negative correlation with unavailable for protein synthesis, this may suggest they
protein digestibility (Marono et al., 2015). Therefore, it could also be unavailable for other typical functions in
 BLACK SOLIDER FLY MEAL FOR BROILER CHICKENS 11
the body, such as for example the lipotropic ability of not viable under the conditions present in this study.
methionine. Methionine works with choline as a methyl However, further research is warranted on BSFLM
donor to export fats from the liver (Kalbande et al., inclusion in lower amounts (≤25% of SBM), as a value-
2009; Saeed et al., 2017). Although liver fat was not ana- added ingredient, which could promote growth similar
lyzed in the present study. It is understood that a methi- to an AGP diet. Further research is also warranted on a
onine deficiency can play a part in the development of comprehensive study of the metabolic and physiological
fatty liver syndrome, a common metabolic condition in effects of BSFLM inclusion, including the effects of high
chickens resulting in fat accumulation in the liver (Qure- levels of BSFLM (≥50%) on organs.
shi et al., 2004). The lack of bioavailable methionine
could have led to the development of fatty liver syn-
drome in this case, causing the increase in liver weight in ACKNOWLEDGMENTS
birds fed higher BSFLM levels. Another factor which This work was supported by Ontario Agri-Food Inno-
could have played a part in the development of fatty vation Alliance, Natural Sciences and Engineering
liver syndrome is the increase in starch contents of the Research Council of Canada, and the Canada First
diets as the BSFLM inclusion level increased. The starch Research Excellence Fund.
content increased due to the increase in corn in the diets.
Corn increased because of reduced SBM as a way to bal-
ance the diets for energy. However, the change in energy DISCLOSURES
source may have influenced the overall composition of
the liver, leading to fatty liver syndrome. The authors declare that the research was conducted
Several other organs became heavier with increasing in the absence of any commercial or financial relation-
BSFLM inclusion. Pancreas weight increased linearly ships that could be construed as a potential conflict of
with the BSFLM on d 24 and 49. It is unclear as to why interest.
pancreas hypertrophy occurred; however, research has
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