The Chemical Content of Fermented Black

Soldier Fly Larvae with Trichoderma viride

as an Alternative Protein Source Feed for Native
Chickens

Kasmira1 , Sulistyawati Ayu Ningrum1 , Sri Purwanti2(B) ,

Wempie Pakiding2 , and Jasmal A. Syamsu2
1 Faculty of Animal Science, Hasanuddin University, Jl. Perintis Kemerdekaan KM. 10,
Makassar 90245, Indonesia
2 Department of Animal Nutrition, Faculty of Animal Science, Universitas Hasanuddin, Jl.
Perintis Kemerdekaan KM. 10, Makassar 90245, Indonesia
sripurwanti@unhas.ac.id

Abstract. Black soldier fly larvae or maggot flour contains high crude fibre which
makes this raw material difficult to digest. High crude fibre content so that the
fermentation process can be carried out as an effort to improve the quality of
maggot flour using the fungus Trichoderma viride microorganism. This study was
conducted to determine the quality of maggot flour fermented with Trichoderma
viride fungus as an alternative protein source feed for native chickens. The research
method used a completely randomized design consisting of 2 factors and 4 repli-
cations so that there were 16 analyzes of the experimental unit. Factor I is the
level of Trichoderma viride (2 and 4%) and the second factor is the time of fer-
mentation (7 and 14 days). The research variables included moisture, ash, crude
fibre, crude protein, and extract ether. The results obtained indicate that the use of
maggot fermented has a significant effect (P < 0.05) on the analysis of moisture,
and extract ether, and in the analysis of ash, crude fibre, and crude protein there
is no effect (P > 0.05). In conclusion, the use of fermented maggot Black soldier
fly flour can be used as a protein source raw material.

Keywords: Black soldier fly · Fermented · native chicken · Trichoderma viride

1 Introduction
Feed ingredients are one of the determinants of the success of an animal’s growth so
that it meets its nutritional needs because it can affect the productivity or growth of
livestock. Several feed ingredients are quite good so that they can be used as potential
raw materials for poultry feed, one of which is maggot flour. The high nutrient content
contained in maggots is the crude protein potential of around 41–42% [1]. Black Soldier
Fly (BSF) (Hermetia illucens, Diptera: Stratiomyidae), is one of the various livestock
that has begun to cultivate Indonesia widely. These flies originate from South America
and then spread to subtropical and tropical regions [2]. Indonesia’s tropical climatic

© The Author(s) 2023

E. Widodo et al. (Eds.): ICESAI 2022, ABSR 28, pp. 438–445, 2023.
https://doi.org/10.2991/978-94-6463-116-6_54
 The Chemical Content of Fermented Black Soldier Fly Larvae 439

conditions are ideal for black soldier fly cultivation. There are many types of insects that
can be exploited as poultry feed ingredients, one of which is black soldier fly. The larvae
of black soldier fly contains 44% dry matter, 42% crude protein, and 35% fat, including
essential amino acids and fatty acids [3]. Black soldier fly is also straightforward to
develop and does not require special equipment. Not only that, but the black soldier fly
cycle is too swift. According to [4], black soldier fly life cycle from egg to adult fly lasts
around 40–43 days, depending on environmental conditions and the feed media given.
The Larvae of black soldier fly contain chitin polysaccharide of about 8.7%, which
can negatively affect the digestibility and utilization of nutrients [5]. The crude fibre is
high enough (exceeding the standard for poultry needs) which is 8.53% and crude fat
is around 27.30% [6] so that the fermentation process can be carried out as an effort to
improve the quality of maggot flour. Reducing the chitin content on organic materials
can be performed by fermentation using specific microorganism that are able to degrade
chitin [7]. The occurrence of fermentation can cause changes in the properties of feed
ingredients due to the breakdown of feed components, The length of fermentation is one
of the factors that greatly affect the length of life and the number of microorganisms that
proliferate. Trichoderma is one of the fungi that can produce chitinase enzymes that can
degrade chitin [8]. Trichoderma spp, is known to produce chitinases, β-13-glucanases,
and proteases. [9] stated that the fungus Trichoderma viride is a producer of complete
cellulose enzymes to hydrolyze cellulose and crystals which causes a decrease in crude
fibre content. [10] who states that Trichoderma viride can increase crude protein and
reduce fibre content. [11] stated that crude fat in fermented products using Trichoderma
viride can also reduce crude fat levels. Based on the quality of post-fermented black
soldier fly maggot flour with BSF, it is suspected that it can be used to improve the
quality of poultry feed ingredients.

2 Materials and Methods

2.1 Making Potato Destro Agar (PDA) Media [12]

Prepare tools and materials to be used. PDA media of as much as 19.5 g was put into
an Erlemmener and dissolved with 500 mL of sterile distilled water, then homogenized
using a centrifuge. Erlenmeyer was closed using a tube cover and then sterilized in an
autoclave at 121ºC for ± 1 h. Then the media was poured into a sterile petri dish and
allowed to solidify. The cup is wrapped so as not to be contaminated and wait for the
media to solidify.

2.2 Transfer of Inoculants to Potato Destro Agar (PDA) Media [13]

Prepare bunsen, ose, and Trichoderma viride isolates. Trichoderma viride isolates were
taken using an ossicle or borer and then transferred to a petri dish containing PDA
media. The petri dish containing the isolate was wrapped so as not to be contaminated
and waited for 1 week until the fungal mycelium grew on the cup.
440 Kasmira et al.

2.3 Propagation of Inoculants in Organic Media (Maize) [14]

Soak the glutinous corn for 1 day. Weigh every 100 g of corn into a heat-resistant plastic
bag. Sterilization of media (corn) by autoclave for ± 1 h. Prepare bunsen, ose and isolate
Trichoderma viride. Take the Trichoderma viride to isolate using ose and then transfer it
to organic media. Re-cover the organic media, and incubate for 1 week until the fungus
grows on the organic media.

2.4 Cellulotic Microbial Activation Using Aerator [9]

Prepare tools and materials to be used, namely Trichoderma viride microbes, molasses,
water, and buckets (containers). Molasses was dissolved in water and then mixed with
Trichoderma viride in container 1 with as much as 2% and container 2 with as much as
4%. Microbes are activated by using an aerator for 24 h and after that, they are ready
to be applied to the maggot flour to be fermented. Fermentation of maggot flour with
Trichoderma viride [15] which maggot flour weighed as much as 1 kg for each treatment
unit. Maggot flour is mixed with activated microbes evenly. Fermentation for treatment
1 for 7 days and fermentation for treatment 2 for 14 days.

2.5 Research Variables

The parameter observed in this study was the content in the proximate analysis of maggot
Black soldier fly flour which was fermented using Trichoderma viride microbes.

2.6 Statistical Analysis

The study used a factorial design based on a completely randomized design (CRD)
consisting of 2 factors and 4 replications, so there were 16 experimental units. The data
obtained were analyzed using analysis of variance by the Factorial Design based on a
Completely Randomized Design (CRD) [16].

3 Results and Discussion

The results of research on the nutrient content with the addition of inoculants with
fermentation time copra meal are presented in Table 1. Table 1 shows the water content
of fermented BSF, at the 2% inoculant level of 9.73 ± 0.66 level and the 4% inoculant
9.73 ± 0.58 and 7 days of fermentation 9.19 ± 0.29 and 14 days of 10.27 ± 0.15.
Analysis of variance showed that the administration of different inoculant levels (2%
and 4%) had no significant effect (P > 0.05) on the water content of fermented maggot
flour, while with different fermentation times (7 days and 14 days) it showed a significant
effect. (P > 0.05) on the water content of fermented maggot flour. Duncan’s test results
showed that the average 7-day fermentation time with a value of 9.19 ± 0.29 was
significantly lower than the 14-day fermentation time with a value of 10.27 ± 0.15,
which indicated that there was an increase in water content of 1.08%. This is because,
at the time of fermentation, microorganisms produce water in their activities, so the
 The Chemical Content of Fermented Black Soldier Fly Larvae 441

Table 1. The average nutritional value of fermented maggot larvae of Trichoderma viride with
different fermentation times

Parameters (%) Number of inoculants (%) Fermentation time (days) Average

7 14
Moisture 2 9.14 ± 0.28 10.32 ± 0.18 9.73 ± 0.66
4 9.23 ± 0.33 10.23 ± 0.12 9.73 ± 0.58
Average 9.19 ± 0.29a 10.27 ± 0.15b 9.73 ± 0.60
Crude fibre 2 9.77 ± 0.39 9.54 ± 0.64 9.65 ± 0.50
4 9.87 ± 0.49 9.40 ± 0.33 9.64 ± 0.46
Average 9.82 ± 0.42 9.47 ± 0.47 9.65 ± 0.47
Crude protein 2% 45.10 ± 1.01 45.13 ± 1.71 45.12 ± 1.31
4% 43.97 ± 1.83 46.83 ± 1.83 45.40 ± 2.14
Average 44.54 ± 1.49 45.98 ± 1.70 45.26 ± 1.72
Crude fat 2% 8.20 ± 0.28a 8.59 ± 0.25a 8.39 ± 0.32a
4% 8.40 ± 0.07a 7.85 ± 0.17b 8.12 ± 0.31b
Average 8.30 ± 0.21 8.22 ± 0.44 8.26 ± 0.34*
Ash 2% 11.68 ± 0.14 11.45 ± 0.11 11.57 ± 0.17
4% 11.58 ± 0.14 11.60 ± 0.09 11.59 ± 0.11
Average 11.63 ± 0.14 11.53 ± 0.12 11.58 ± 0.14
a, b : Different superscripts in the same row and column show significant differences (P<0.05)

longer the fermentation, the more water is produced. In line with the opinion of [17]
who states that the increase in water content in the fermentation process is caused by
the increasing number of mushrooms so the water content increases. This is because in
the fermentation process there is a breakdown of glucose into carbon dioxide, thereby
increasing the water content of the fermented material.
Table 1 shows the crude fiber test values with the average fiber content of fermented
BSF larvae, at the 2% inoculant level of 9.65 ± 0.50, the 4% inoculant level of 9.64 ±
0.46% and the 7-day fermentation time of 9.82 ± 0.42% and 14 days 9.47 ± 0.47%.
Analysis of variance showed that the application of different inoculant levels and fer-
mentation time gave different mean values of crude fiber. The 2% inoculant level gave
a higher value than the 4% level so there was an increase of about 0.01% the 7-day
fermentation period was higher than 14 days and there was a decrease of 0.35%. Statis-
tically, inoculant level and fermentation time had no significant effect (P > 0.05) on the
crude fiber value of fermented maggot flour. According to [18], the higher the microbial
population in fermented feed, the lower the crude fiber, this is because the inoculant
activity of lignocellulolytic bacteria degrades the fiber faster.
Table 1 shows the average value of crude protein content of fermented BSF larvae
at the 2% inoculant level of 45.12 ± 1.3% and the 4% inoculant level of 45.40 ± 2.14a
and at the 7-day fermentation time of 44.54 ± 1.49% and at the 14-day fermentation
442 Kasmira et al.

9
8.5

Extract Ether (%)

8
7.5 2%
7 4%
7 14

Fermentation periode (days)

Fig. 1. Decreasing crude fat content with different levels of inoculants and fermentation time

time of 45.98 ± 1.70%. Analysis of variance showed that the administration of different
levels of inoculants showed different levels of crude protein. As seen in the addition
of a 2% inoculant level, crude protein content was lower than the 4% level and there
was an increase of about 1.44%. Apart from the inoculant level, the 7-day fermentation
time was lower than 14 days and there was an increase of 0.28%. However, statistically
inoculant level and fermentation time did not significantly affect (P > 0.05) the crude
protein value of fermented maggot flour. Research conducted by [19] shows that maggot’s
protein content is relatively high. Maggot in dry form contains 41–42% crude protein
[20] in their research on the effect of the fermentation time of a mixture of Trichoderma
reesei and Aspergillus niger on the protein and crude fiber content of year dregs that the
longer the incubation time, the higher the protein content, the increase in protein content
was due to the addition of protein given by microbial cells derived from their growth to
produce single cell products. The statement [21] in their research is that Trichoderma
viride microbes can increase crude fiber protein, this is because Trichoderma viride can
utilize organic materials contained in the substrate to be broken down and convert them
into increased protein content.
Table 1 shows the proximate test values for the crude fat content of fermented BSF
larvae at the 2% inoculant level of 8.39 ± 0.32% and the 4% inoculant level of 8.12
± 0.31%, while the 7-day fermentation period was 8.30 ± 0.21% and the fermentation
time was 14 days. of 8.22 ± 0.44%. Analysis of variance showed that the administration
of the 2% inoculant level was higher than the 4% level so there was a decrease of
about 0.27% and the 7-day fermentation period was higher than the 14-day fermentation
period and there was an increase of 0.08%. The results of Duncan’s test showed that
the inoculant level and fermentation time had a significant effect (P > 0.05) on the
crude fat value of fermented maggot flour. [22] states that the longer fermentation time
can reduce crude fat content because the microbial activity is more active in degrading
complex compounds into simple compounds to facilitate fat degradation from fermented
materials. Analysis of variance showed that the interaction between the inoculant level
and different fermentation times had a significant effect (P < 0.01) on the crude fat value
of BSF maggot larvae flour. In addition, there is an interaction between the administration
of the inoculant level and the duration of fermentation. Table 1 shows the proximate test
values for the ash of fermented BSF larvae at the 2% inoculant level of 11.57 ± 0.17%
and the 4% inoculant level of 11.59 ± 0.11%, while the 7-day fermentation period was
 The Chemical Content of Fermented Black Soldier Fly Larvae 443

11.63 ± 0.14% and the fermentation time was 14 days. of 11.53 ± 0.12%. It increases
ash content due to the decreased organic matter after fermentation. This is presumably
due to the activity of Trichoderma viride extracellular enzymes that degrade organic
matter of BSF larvae especially fat content that causes shrinkage of organic matter. The
decrease in organic matter resulted in an increase in the proportion of ash content. This
is in line with the studies of [23] that fermentation increases the ash content. In Fig. 1, it
was found that the administration of 2% and 4% inoculant levels resulted in a decrease in
crude fat values. At 7 days of fermentation, the level of inoculant administration of 2%
approached the same point while the level of salt administration of 4% was at a higher
point which indicated that the level of inoculant administration of 2% did not reduce fat
content. The decrease in crude fat value occurred along with the addition of inoculant
levels and fermentation time.

4 Conclusions

Concluded that Trichoderma viride fermented BSF maggot flour decreased crude fat
content by 0.27% in 2% and 4% inoculants and an increase in water content at 7 and
14 days of fermentation by 1.08% and can be used as raw material for protein source
feed.

Acknowledgment. The author would like to express my sincere thanks to the Ministry of
Research, Technology and Higher Education in providing BPPDN scholarships in our studies and
research, as well as the staff of the biotechnology and waste valorization laboratory at Hasanuddin
University who provided laboratory equipment and research facilities during the experiment.

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