sustainability
Article
Fish Food Production Using Agro-Industrial Waste Enhanced
with Spirulina sp.
Margarita Ramírez-Carmona
Citation: Ramírez-Carmona, M.;
Rendón-Castrillón, L.;
Ocampo-López, C.; Sánchez-Osorno,
D. Fish Food Production Using
Agro-Industrial Waste Enhanced
with Spirulina sp. Sustainability 2022,
14, 6059. https://doi.org/10.3390/
su14106059
Academic Editor: Mark Flaherty
Received: 6 April 2022
Accepted: 11 May 2022
Published: 17 May 2022
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4.0/).
Sustainability 2022, 14, 6059. https://doi.org/10.3390/su14106059
, Leidy Rendón-Castrillón, Carlos Ocampo-López * and Diego Sánchez-Osorno
Facultad de Ingeniería Química, Centro de Estudios y de Investigación en Biotecnología (CIBIOT),
Universidad Pontificia Bolivariana, Circular 1A No. 70-01, Building 11, Medellín 050031, Colombia;
margarita.ramirez@upb.edu.co (M.R.-C.); leidy.rendon@upb.edu.co (L.R.-C.);
diego.sanchezo@upb.edu.co (D.S.-O.)
* Correspondence: carlos.ocampo@upb.edu.co
Abstract: The supply of animal feed is one of the main concerns of producers in the aquaculture
industry, including aspects such as the cost of fish flour and its nutritional balance. The aim of
this study was the preparation of a pellet-type fish food using powdered Spirulina sp. cultivated
as a protein source supplemented with agro-industrial waste, and its evaluation to comply with
the necessary parameters for the elaboration of extruded pellets. Spirulina sp. was cultivated in a
photobioreactor at a volume of 50 L, separated by decantation and dried. The proximal character-
ization was 6.79% ± 0.05 moisture, 6.93% ± 0.01 ash, 66.88% ± 0.33 protein, and 5.50% ± 0.26 fat.
Subsequently, flours were prepared using cassava leaves, gliricidia leaves, and rice husks. The results
for the cohesion showed that the flours obtained to comply with the necessary parameters for the
elaboration of extruded food. The fish feed was prepared in pellet form using the formulation for
fattening Tilapia: Spirulina sp. (20%), cassava leaf flour (50%), gliricidia leaf flour (20%) and flour of
rice husk (10%). Floatation analysis showed that 60% of the pellets floated for more than 40 min, and
80% retained their shape for 4 h. The results show that the obtained product can be used as fish feed,
due to the lowest disintegration, together with its great capacity for water absorption and especially,
its greater flotage due to the expansion effect, are physic characteristic determinants so that the fish
has more time to consume extruded diets and avoid losses.
Keywords: cassava leaves; gliricidia leaves; rice husk; residual biomass; aquaculture; agricultural
biomass
1. Introduction
Agricultural production increased more than three times in the last 50 years due to
the expansion of land for agricultural use, the technological contribution of the green
revolution that influenced productivity, and the accelerated growth of the population [1].
This sector produces an average of 23.7 million tons of food per day worldwide [1].
Likewise, the agricultural waste production is estimated at 998 million tons per year,
made up of animal waste, food processing waste, crop waste such as corn stalks, sugarcane
bagasse, discarded fruit and vegetables, and pruning, among others. Organic waste can
represent up to 80 percent of the total solid waste generated on any farm [2].
The growth of world agricultural production creates greater pressure on the environ-
ment, causing negative impacts on soil, air, and water resources [1,3]. Furthermore, it can
lead to the loss and degradation of natural ecosystems [4].
In general, agriculture is responsible for 21% of greenhouse gas emissions [1]. Recently,
this situation has promoted a more sustainable development model, which implies relevant
changes in current agricultural production systems [5]. For example, use of agricultural
residues to fertilize soils and contribute to the dispersion of chemical and biological pollu-
tants in terrestrial environments [6], the use of residues for cooking and heating [7], as well
as the production of biogas [8] and the production of alternative protein for animal feed [9].
https://www.mdpi.com/journal/sustainability
Sustainability 2022, 14, 6059 2 of 17

These processes are classified as waste recovery waste into more suitable products,
with opportunities to apply industrial symbiosis in waste recovery. Consequently, it has
potential to provide economic, social, and environmental benefits. These strategies support
the development of a circular economy in the food sector by closing the loop and using
waste as a resource, an approach encouraged by recent policies in Europe [10].
Circular economy is defined as the conservation of the products value, materials, and
resources in the economy for a long period with the reduction of waste generation. The
circular economy aims to improve the efficiency of resource use and the recovery of waste
materials to reduce the emission of additional fossil carbon during manufacturing and
extraction processes. In this sense, the sustainable and synergistic use of resources should
be encouraged with bioeconomy instead of increasing pressure on resources [5].
One of the practices that represent low use of agricultural land is the production
of animal feed using new protein sources such as insect meal, micro, and macroalgae,
protein-rich by-products of the food industry, and unicellular protein [9], as well as taking
advantage of poultry and papaya processing waste used as alternative proteins to replace
fishmeal [11].
In aquaculture, the supply of animal feed is one of the main concerns of producers in
this industry [12] due to limited resources such as land, water, and energy [13], where 560
Mha, or about 40% of the world’s agricultural land in the production of animal feed [14].
Feeding costs and quality are the main constraints to the emergence of aquaculture in
developing countries. Fish feeding represents 60–75% of the total cost of fish production.
The high cost of food is due to the high cost of fish flour, the main ingredient in commercial
food formulation [15].
The market drives substitute marine ingredients to different plant-based ingredients,
and novel food ingredients are driven by price, availability, and legislation [16]. Alterna-
tive feeding solutions such as the use of insects can have unexpected side effects when
implemented and expanded to industrial production [12,13].
The use of fish flour decreases every year, reaching an approximate consumption of
141,000 tons in 2017, of which 69% was used for aquaculture in competition with food
security [17]. The use of microalgae as a fish feed or food supplement could reduce the
burden on fish flour-based aquaculture; and reduce the gap between fish production and
demand. Similarly, when microalgae are mixed with animal feed for the required nutrient
portion, the requirement of grains and millet could be reduced [18].
Microalgae have several attractive characteristics for large-scale sustainable produc-
tion, such as high biomass yields per unit area and grow ability on non-arable land using
non-potable water or even saltwater [19].
The Spirulina sp. microalgae is a potential source of nutrients as food supplementation
in aquaculture due to its high content of protein and absorbable compounds [20], as well
as antioxidant properties, improving growth, feeding efficiency, and the physiological
response to disease in various fish species at low production cost [21].
The production of fish feed using locally available ingredients at a low cost is crucial
for the development and sustainability of aquaculture [15,22].
In this study, a pelletized fish food production was developed using agro-industrial
wastes composed of cassava leaf flour, gliricidia leaves, and rice husk, enhanced with
powder Spirulina sp. as a protein source. A set of photobioreactors to produce Spirulina sp.
were installed in rural areas of the Department of Antioquia, Colombia, and a cooperative
production model was implemented with farmers to guarantee the long-term sustainability
of the initiative.

2. Materials and Methods

2.1. Raw Materials for the Processing of Fish Food
The raw materials used to prepare fish food were: Spirulina sp. obtained from CIBIOT
lad at Universidad Pontificia Bolivariana, cassava leaves (CL) and gliricidia leaves (GL)
Sustainability 2022, 14, 6059 3 of 17

from the field -San Carlos Antioquia- Colombia, and rice husk (RH) from the Plaza May-
orista de Medellín.

2.2. Spirulina sp. Production

The Spirulina sp. culture was developed in a 50 L photobioreactor, bubbling system
and, an inoculum concentration equivalent to 20% at room temperature.
The culture medium used to produce this Spirulina sp. was the modified Zarrouk
medium, as shown in Table 1. The trace metal solution was composed of Boric acid
23.13 mol·m−3 , Manganese chloride tetrahydrate 4.55 mol·m−3 , Zinc sulfate heptahydrate
0.52 mol·m−3 , Copper sulfate pentahydrate 0.16 mol·m−3 , Sodium molybdate dihydrate
0.05 mol·m−3 .

Table 1. Modified Zarrouk medium.

Compound Concentration (mol·m−3 )

NaHCO3 200
K2 HPO4 3.67
NaNO3 29.41
K2 SO4 5.73
NaCl 17.11
MgSO4 7H2 O 2.62
CaCl2 2H2 O 0.27
FeSO4 7H2 O 0.04
EDTA 200

The production of Spirulina sp. was monitored every 4 days in a physical way employ-
ing photographic recording, microscopic review, and determination of the optical density
by the spectrophotometric method between 400 and 700 nm.
Subsequently, the biomass was separated by decantation and subjected to convection
oven drying for 24 h at 65 ◦ C to avoid protein denaturation and subsequent maceration of
the biomass on a dry basis using a mortar. The algal biomass on a dry basis, the percentage
of moisture, protein, ash, and fat was determined. To determine the moisture content, the
AOAC 964.22 method was used, the wet sample was taken to the drying oven at 105 ◦ C
for 24 h. The samples were analyzed in duplicate, and the results were expressed as a
percentage of moisture.
Ash was determined by calcining the samples in an electric muffle (TERMOLYNE,
model 1300), at a temperature of 550 ◦ C for 4 h (AOAC 1990). The results obtained were
expressed as a percentage of ash.
The protein percentage was determined indirectly using elemental analysis (CHON)
(Carbon: AOAC 949.14, Hydrogen: AOAC 949.14, Nitrogen: AOA 984.13). The analysis
was performed using Thermo Scientific FLASH 2000 Analyzer, Organic Elemental Analyzer.
For the determination of fat content, the modified Blight and Dyer method, adopted
by AOAC in 1990 was used. The samples were analyzed in duplicate, and the results were
expressed as a percentage of fat.

2.3. Manufacture of Flours from Agro-Industrial Waste

To produce flour from the raw materials (CL, GL, and RH), a milling process was
carried out using a TE-651/2 Moihno mill from Rotor TECNAL. Each raw material was
separated three times using mesh sieves numbers 10, 20, and 30, respectively. Subsequently,
screening was carried out using sieves 12, 16, 20, 30 and 40, finally, the particle size
distribution was determined according to ASTM E 11-61, and particle size less than 420 µm
was selected (Sieve No. 40).
Both cassava and gliricidia leaves were previously subjected to a washing process
with acetic acid of 200 ppm concentration for 10 min, then they were subjected to a drying
process at 60 ◦ C by oven convection for 48 h.
Sustainability 2022, 14, 6059 4 of 17

The flours were characterized by performing a proximal analysis in duplicate: moisture

according to the AOAC 964.22 method, ash by calcination, the percentage of protein
indirectly by elemental analysis using a Thermo Scientific FLASH 2000 equipment, Organic
Elemental Analyzer and fat by the Blight method, and modified Dyer, adopted by AOAC
in 1990.
The particle size of the flours was determined through image analysis using a stere-
omicroscope (transmitted light) with a 4.0× Nikon SMZ (500 objective) and a Nikon DS-Vi1
digital camera connected to a USB interface adapted to the system. The capture was made
with a constant focal length and standardized light conditions. Images were analyzed with
Image J software (V2.1, Optika Vision Lite, Ponteranica, Italy).
The Cohesion Index (CI) was determined using a 48 mm diameter and 10 mm high
rotary blade [23,24] while the flow characteristics of the flours were evaluated using a TA
texturometer. X T PLUS Texture Analyzer (Stable Micro Systems, Medellín, CO, USA) [23].

2.4. Processing of Food for Fish

The concentrate was prepared using the nutritional requirements of Red Tilapia. Three
diets were formulated and manufactured aiming to have diets with similar protein contents
that could be used at different times of the year where there is variation in the availability
of raw materials to produce fish food. A balanced feed with a net protein content of
about 35% was formulated for the formulations. The ingredients of the formulation were:
Spirulina sp. powder, cassava leaf flour, gliricidia leaf flour, and rice husk flour, according
to the formulation shown in Table 2.

Table 2. Percentages of flours used in the formulation of fish feed (mass percent, dry basis).

Flours (%)
Formulation Spirulina sp. Cassava Leaves Gliricidia Leaves Rice Husk
1 20 40 20 20
2 35 35 20 10
3 20 50 20 10

The prepared homogenous blend of flours was extruded in an extruder (Haake Polylab
OS). Experiments were performed at a screw speed of 300 rpm and feed screw speed of
175 rpm. The barrel temperature was 100 ◦ C. The configurations of extruder are same as
discussed by Seth and Rajamanickam [25].
The extrusion process for the formation of the pellets was carried out by mixing the
flours until homogeneity was achieved according to the formulations shown in Table 2, then
water was added during the stirring process until reaching 30% moisture in the mixture
and then they were processed in a 150 mm long (L) and 10 mm diameter (D) single worm
screw extruder with a 15:1 L/D ratio. The compression ratio of the screw was 5:1 and the
diameter of the hole of the nozzle used was 5 mm, temperature 127.5 ◦ C, speed 160 (rpm),
and moisture 30%. The extruded cords were cut into pellets, with a length of approximately
5 mm. These were subsequently dried in a BINDER oven at 60 ◦ C until reaching a moisture
of 10 ± 0.5%. Feed pellets were not coated with fish oil, as usually performed during
large-scale production as we wanted to exclude the impact of a coating step on the quality
of the fish feed. The pellets were determined to have floating and stability in water.
For the floating test 10 pellets were taken and deposited in a beaker with 400 mL of
water at 25 ◦ C, every 5 min the pellets that were kept on the surface were counted and the
percentage of floating was determined by the method proposed by Arroyo [26].
For the stability test in water, 8 g sample of pellets was deposited in a beaker with
500 mL of distilled water at 25 ◦ C, without stirring, then this mixture was passed through
a # 8 mesh sieve allowing the solid particles to be retained. The mesh along with the
sample was dried using a Thermolab convection dryer for 24 h at 60 ◦ C. The stability of the
 For the floating test 10 pellets were taken and deposited in a beaker with 400 mL of
water at 25 °C, every 5 min the pellets that were kept on the surface were counted and the
percentage of floating was determined by the method proposed by Arroyo [26].
For the stability test in water, 8 g sample of pellets was deposited in a beaker with
500 mL of distilled water at 25 °C, without stirring, then this mixture was passed through
Sustainability 2022, 14, 6059 5 of 17
a # 8 mesh sieve allowing the solid particles to be retained. The mesh along with the sam-
ple was dried using a Thermolab convection dryer for 24 h at 60 °C. The stability of the
pellet in terms of dry matter retention (DMR) was calculated with the weight of the dry
pellet inusing
sample terms of dry matter retention (DMR) was calculated with the weight of the dry
sample using
% DMR PWbi − PWai∗ 100, (1)
% DMR = ∗ 100, (1)
PWbi
where, DMR: dry matter retention (%); PWbi: pellet weight before immersion (kg); PWai,
where,weight
pellet DMR:after
dry matter retention
immersion (kg). (%); PWbi : pellet weight before immersion (kg); PWai ,
pellet weight after immersion (kg).
3. Results and Discussion
3. Results and Discussion
3.1.
3.1. Raw
Raw Materials
Materials forfor the
the Processing
Processing of of Fish
Fish Food
Food
3.1.1. Spirulina
3.1.1. Spirulina sp. sp.
In
In this
this research,
research, Spirulina
Spirulina sp.sp. was
was cultivated
cultivatedto to be
be used
used asas aa source
source of of protein
protein inin the
the
elaboration of fish feed, between
elaboration of fish feed, between 51 and 51 and 71% by dry weight, essential and nonessential
nonessential
amino
amino acids.
acids. TheThe protein
protein content
content of of Spirulina
Spirulina sp.sp. isis higher
higher when
when compared
compared to to unicellular
unicellular
algae
algae andand cyanobacteria,
cyanobacteria, they they also
also contain
contain fourfour times
times more absorbable
absorbable compounds
compounds than than
protein in meat [20,27].
Spirulina
Spirulina sp. sp. as
as anan additive
additive in in animal
animal feedfeed improves
improves growth,
growth, feeding
feeding efficiency,
efficiency, and
and
the physiological
the physiological response
response to to disease
disease in in various
various fishfish species
species [28,29].
[28,29]. Also,
Also, itit has
has received
received
certification by the
certification the Food
Food andand Drug
Drug Administration
Administration (FDA) (FDA) asasaafood
foodor ordietary
dietarysupplement.
supplement.
Studies by Nevien
Studies Nevienet etal.
al.[21]
[21]evaluated
evaluatedthe theeffect
effectof of
dietary supplementation
dietary supplementation for Tilapia
for Ti-
lapia Spirulina
usingusing platensis
Spirulina showing
platensis improvements
showing improvements in markers
in markersof liver andand
of liver kidney
kidneydamage
dam-
in fish
age exposed
in fish exposedto subacute
to subacutediazinon toxicity
diazinon (DZN).
toxicity Besides,
(DZN). biochemical
Besides, markers
biochemical were
markers
improved
were through
improved the increase
through of albumin
the increase of albumin andand globulins
globulinswith a significant
with a significant decrease
decrease in
glucose and cholesterol. Furthermore, there was an improvement
in glucose and cholesterol. Furthermore, there was an improvement in the liver, kidneys, in the liver, kidneys, and
gills gills
and afterafter
beingbeing
exposed to DZN
exposed wherewhere
to DZN a significant increase
a significant in tissue
increase inantioxidant activity
tissue antioxidant
was observed with a significant decrease in lipid peroxidation
activity was observed with a significant decrease in lipid peroxidation levels [21]. levels [21].
Spirulina platensis
Spirulina platensis was
was also
also evaluated
evaluated as as aa 1%
1% dietary
dietary supplement
supplement in in common
common carp carp
(Cyprinus carpio L.)
(Cyprinus carpio L.) induced
induced by by oxidative
oxidative stress
stress and
and Atrazine
Atrazine inflammation
inflammation (ATZ) (ATZ) [30].
[30]. In
In
Figure 1, the cultivation and adaptation of Spirulina sp. Cyanobacteria
Figure 1, the cultivation and adaptation of Spirulina sp. Cyanobacteria in a 50 L bioreactor in a 50 L bioreactor
are shown,
are shown, using
using an an inoculum
inoculum concentration
concentration equivalent
equivalent to to 20%.
20%.

Figure 1. Culture of Spirulina sp. in a 50 L bioreactor.

One of the main nutrients required by Spirulina sp., for its growth is the source
of Carbon, one of the direct sources in controlled environments, is sodium bicarbonate,
commercial or analytical grade, which not only acts as a medium that provides carbon to
the cultivation but also regulates its alkaline conditions; relevant factor for maintaining the
sterility of the medium and the pH in a range of 9 to 11 [31,32].
Similarly, as evidenced in the culture medium, this cyanobacterium requires a source
of Nitrogen in the medium, generally supplied by NaNO3 , which is essential for its growth.
This source is also required for the synthesis of amino acids that structure proteins and
 Oneof
One ofthe
themain
mainnutrients
nutrientsrequired
requiredby bySpirulina
Spirulinasp.,sp.,for
forits
itsgrowth
growthisisthethesource
sourceof of
Carbon, one of the direct sources in controlled environments, is
Carbon, one of the direct sources in controlled environments, is sodium bicarbonate, com- sodium bicarbonate, com-
mercialor
mercial oranalytical
analyticalgrade,
grade,which
whichnot notonly
onlyacts
actsas
asaamedium
mediumthat thatprovides
providescarbon
carbontotothe the
cultivation but also regulates its alkaline conditions; relevant factor
cultivation but also regulates its alkaline conditions; relevant factor for maintaining the for maintaining the
sterility of the medium and the pH in a range
sterility of the medium and the pH in a range of 9 to 11 [31,32]. of 9 to 11 [31,32].
Sustainability 2022, 14, 6059 6 of 17
Similarly,as
Similarly, asevidenced
evidencedininthe theculture
culturemedium,
medium,this thiscyanobacterium
cyanobacteriumrequiresrequiresaasource
source
ofof Nitrogen
Nitrogen inin the the medium,
medium, generally
generally supplied
supplied by by NaNO
NaNO3,3, which
which isis essential
essential for for its
its
growth. This source is also required for the synthesis of amino
growth. This source is also required for the synthesis of amino acids that structure pro- acids that structure pro-
other cellular
teinsand
andother components
other such as phycocyanin,
cellularcomponents
components suchas the main the
asphycocyanin,
phycocyanin, pigment of phycobiliproteins
mainpigment
pigment ofphycobili-
phycobili-
teins cellular such the main of
that, in addition
proteins that, in to having to
addition a photosynthetic
having a primary function,
photosynthetic primary help the reserve
function, help the ofreserve
cellularof
proteins that, in addition to having a photosynthetic primary function, help the reserve of
nitrogen [32–34].
cellularnitrogen
nitrogen[32–34].
[32–34].
cellular
The
The cultivation
cultivation ofofSpirulina
Spirulinasp.sp.
requires
requires a source such as phosphorus, which is essential
The cultivation of Spirulina sp. requires aasource
source such
such asphosphorus,
as phosphorus, which
which isisessen-
essen-
intial
theinperformance
the performance of this ofcyanobacterium
this cyanobacterium because it maintains
because it the
maintainsproduction
the of
production biomass
ofbi-
bi-
tial in the performance of this cyanobacterium because it maintains the production of
inomass
the culture;
in the the form the
culture; thatform
the Phosphorus
that the of Spirulina
Phosphorus ofsp. acquires
Spirulina isacquires
sp. through is inorganic
through
omass in the culture; the form that the2−Phosphorus of Spirulina sp. acquires is through
phosphates such as H2such
inorganicphosphates
phosphates PO4 −asorHHPO . In Figure 2, it is shown the follow-up to the
2PO4−4or HPO42−. In Figure 2, it is shown the follow-up to
inorganic
Spirulina sp. culture, to
such
show
as H
the
2PO4− or HPO42−. In Figure 2, it is shown the follow-up to
adaptation and growth of theofculture.
theSpirulina
the Spirulinasp. sp.culture,
culture,totoshowshowthe theadaptation
adaptation andgrowth
and growth theculture.
of the culture.

Figure2.2.Monitoring
Figure MonitoringofofSpirulina
Spirulinasp.
sp.by
byoptical
opticaldensity
densitybetween
between400
400and
and700
700nm.
nm.
Figure 2. Spirulina sp. 400 and 700 nm.

InInFigure
Figure2,
2,2,the
theadaptation
the adaptationofofSpirulina
adaptation Spirulinasp.
sp.to
tothe
theculture
cultureconditions
conditionsisisevidenced,
evidenced,in- in-
cluding
cluding factors
factors such
such asas
pH,pH, light
light intensity,
intensity, presence
presence of of contaminants,
contaminants, temperature,
temperature,
cluding factors such as pH, light intensity, presence of contaminants, temperature, pres- pres-
presence
ence
of
ence ofofbicarbonate
bicarbonate
bicarbonate ions,nitrogen
ions, ions,
nitrogennitrogen
source, source, type
type type
source, of ofbioreactor,
bioreactor,
bioreactor,
of initial
initialinitial
biomass biomass concentration
concentration
biomass and
concentration
and population
population
and population density density [35–37].
[35–37].
density [35–37].
InInthe
In thesame
the sameway
same wayin
way inFigure
in Figure3,
Figure 3,3,the
thefollow-up
the follow-upmade
follow-up madeto
made totothe
theculture
the cultureof
culture ofofSpirulina
Spirulinasp.,
Spirulina sp.,under
sp., under
under
themicroscope
the
the microscopeMotic
microscope Moticbrand
Motic brandbinocular-1100100401401
brand binocular-1100100401401ba210led,
binocular-1100100401401 ba210led,is
ba210led, isisevidenced,
evidenced,using
evidenced, usingthe
using the
the
10X/0.25
10X/0.25 (WD
(WD 17.4
17.4 mm)
mm)
10X/0.25 (WD 17.4 mm) objective. objective.
objective.

Figure 3. Monitoring of Spirulina sp. under a microscope Motic brand binocular-1100100401401

ba210led, using the 10X/0.25 (WD 17.4 mm) objective.

In Figure 3, it is observed that the morphology of Spirulina sp., has a spiral-shaped

structure, composed of multicellular cylindrical trichomes, which are arranged in the form
of an open helix, along its length, where environmental factors, such as temperature and
physical and chemical conditions, affect the helix geometry [31,36].
After the production of the Spirulina sp. biomass, it was separated by decantation as
shown in Figure 4.
 In Figure 3, it is observed that the morphology of Spirulina sp., has a spiral-shaped
structure, composed of multicellular cylindrical trichomes, which are arranged in the form
of an open helix, along its length, where environmental factors, such as temperature and
physical and chemical conditions, affect the helix geometry [31,36].
Sustainability 2022, 14, 6059 7 of 17
After the production of the Spirulina sp. biomass, it was separated by decantation as
shown in Figure 4.

Figure 4.
Figure 4. Decantation process
process of
of Spirulina
Spirulina sp.
sp.

In
In Figure 4, 4, the
thedecanting
decantingprocess
processofofthe
the Spirulina
Spirulina sp.sp. culture
culture is evidenced,
is evidenced, and
and later
later this subjected
this was was subjected to a convection
to a convection oven drying
oven drying processprocess
for 24 for
h at24a h at a temperature
temperature of
of 65 °C,
to ◦avoid
65 C, to protein
avoid protein denaturation,
denaturation, where studies
where studies carried carried out by Pantoja
out by Pantoja et al.,
et al., show show
that the
that ◦ C and the exposure time
best the best temperature
temperature for ovenfor ovenof
drying drying
algal of algal biomass
biomass is 52 °C is
and52the exposure time of 3 to 4
of 3 toas4 days, ◦
days, it is a as it is a heat-labile
heat-labile product,
product, which which
starts at 65starts
°C toatdecompose
65 C to decompose into its
into its nutritional
nutritional properties, as proteins are denatured and great content of minerals
properties, as proteins are denatured and great content of minerals and vitamins is lost and vitamins
is lost [38].
[38].
Subsequently,
Subsequently, upon upon obtaining
obtainingthe thebiomass
biomassonona drya dry basis,
basis, thisthis
waswas characterized,
characterized, ob-
obtaining a percentage
taining a percentage of 6.79%
of 6.79% ± 0.05
± 0.05 of moisture,
of moisture, 6.93% ± 0.01±of0.01
6.93% ash,of66.88% ± 0.33 ±
ash, 66.88% of 0.33
pro-
of
teinprotein
and 5.50% and 5.50%
± 0.26 of ± fat.
0.26The
of fat.
aboveThe above coincides
coincides with studieswith studies
carried outcarried
by Lara out
et by
al.
Lara et al. [39], who present this cyanobacterium with great potential
[39], who present this cyanobacterium with great potential in protein biosynthesis and in protein biosynthesis
and growth.
growth. Furthermore,
Furthermore, the results
the results show show that cultivated
that cultivated Spirulina
Spirulina sp. can sp. can be
be used used for
as food as
food for fish, especially Tilapia. These require a source of protein
fish, especially Tilapia. These require a source of protein of 35–45% and fat of 10–15% of 35–45% and fat of
10–15%
[40,41]. [40,41].
The
The results
results of the Spirulina
of the Spirulina sp.
sp. cultivated
cultivated in
in this
this investigation
investigation present
present protein
protein values
values
of 66.88% ±
of 66.88% 0.33, values
± 0.33, values that
that are
are compared
compared with the Arthrospira
with the Arthrospira platensis
platensis species
species that
that have
have
up
up toto 70%.
70%. The The rest
rest of
of the
the algae
algae species
species average
average around
around 40%40% protein
protein content,
content, which
which is is
relatively high compared to other plant sources such as soy (38%), rice (~10%), pea (2.8%)
relatively high compared to other plant sources such as soy (38%), rice (~10%), pea (2.8%)
or even animal sources like milk (4%) or eggs (13%). Algae are not only high in protein, but
or even animal sources like milk (4%) or eggs (13%). Algae are not only high in protein,
that protein’s composition is much richer in essential amino acids compared to common
but that protein’s composition is much richer in essential amino acids compared to com-
plant proteins [19,27,42].
mon plant proteins [19,27,42].
Microalgae can be produced in large volumes at relatively low costs, and reportedly
Microalgae can be produced in large volumes at relatively low costs, and reportedly
can produce 7000 to 10,000 kg per hectare per year, making it a potential source of protein
can produce 7000 to 10,000 kg per hectare per year, making it a potential source of protein
for aquaculture [37,43].
for aquaculture [37,43].
3.1.2. Flours Produced from Agro-Industrial Waste
3.1.2. Flours Produced from Agro-Industrial Waste
The raw materials for the elaboration of the fish concentrate were selected according
to theThe raw materials
protein content, the fornutritional
the elaboration of the
quality, thefish concentrate
availability were
of the rawselected according
material and the
to the
low protein
costs content,
tending towardsthe the
nutritional quality,
sustainability ofthe
fishavailability
farming. of the raw material and the
low costs tending
Therefore, towards
in this the sustainability
research of fish farming.
agricultural residual biomasses were used: cassava leaves
Therefore, in this research agricultural residual
(Manihot esculenta), gliricidia leaves (Gliricidia sepium), biomasses were
and rice used:
husk cassava
(Oryza leaves
sativa) to
(Manihot esculenta), gliricidia leaves (Gliricidia sepium),
produce flours as raw materials in the production of fish food. and rice husk (Oryza sativa) to pro-
duceTheflours as raw
market hasmaterials in the production
been promoting of fishoffood.
the substitution marine ingredients for a variety of
ingredients of plant origin with positive socio-economic and environmental impacts [16,18]
to improve local fish production, as it remains marginal with low yields [15].
The residual biomass generated from cassava cultivation on an annual average is
13.39 Ton/Ha, while for gliricidia cultivation it is an annual average of 2.25 Ton/Ha [44]
and 1.6 Ton/Ha of rice husk, what guarantees local raw material. Today, finding sustainable
raw materials is a major obstacle and has become a priority among stakeholders for fish
feed production [13].
Cassava cultivation is distributed in diverse tropical countries with an estimated
annual production of 291.9 million tons. The crop is the most important source of carbohy-
Sustainability 2022, 14, 6059 8 of 17

drates in producing countries. Both the roots and the leaves of the cassava can be used as
food. Cassava is the most important source of carbohydrates in producing countries and
can additionally contribute to the supply of proteins, micronutrients, and minerals [8].
Cassava is also the third most important source of calories in the tropics and the
sixth most important crop in annual global production. Cassava crop is relevant for small
farmers and large-scale plantations because of its low nutrient requirement, the ability to
tolerate dry conditions, and the easy propagation at a low cost. The reliance on this crop is
expected to increase as the global climate changes [45].
Although cassava leaves are an important source of carbohydrates and contribute
to the supply of proteins, micronutrients and minerals, they must be detoxified before
being used in animal or human nutrition due to the cyanogenic components that vary
in concentration depending on the variety, genetics and growth stage [8]. Cyanogenic
compounds release hydrogen cyanide. The degree of toxicity is altered by plant breeding,
agricultural practice, environmental conditions, and food preparation methods [45].
Cyanide, in cassava leaves, can be found as free cyanide (approx. 10% lotaustraline)
and as bound cyanide (approx. 90% linamarin), the latter being the contributor of the
cyanide ion for the formation of hydrocyanic acid as free cyanide.
The crushing process in cassava leaves allows the enzyme linamarase to act on limarin
due to the increase in the surface area obtained with the breakdown of cell walls during
crushing. Increasing the proportion of free cyanide to ranges between 30–40%. Then, the
free cyanide is eliminated during the dehydration process due to the favor of the hydrolytic
reaction with long drying times and low temperatures.
Table 3 shows the characterization of raw materials used in this study, in mass percentage.

Table 3. Characterization of raw materials, in mass percentage.

Raw Materials % Moisture % Ashes % Protein % Fat

Cassava leaf flour 10.23 ± 0.15 8.91 ± 0.0 28.33 ± 0.15 4.42 ± 0.19
Gliricidia leaf flour 9.15 ± 0.14 5.94 ± 0.0 25.63 ± 0.09 3.43 ± 0.52
Rice husk flour 7.32 ± 0.18 16.50 ± 0.01 7.17 ± 0.02 0.61 ± 0.10

The results of Table 3 show that the moisture percentages of the raw materials were
less than 13%, this being considered a good quality factor framed in the codex standard
176-1989 and codex standard 193-1995 [46].
Gliricidia is a plant that spreads rapidly. In Brazil, Gliricidia has often considered
an invasive or weedy species and is employed as a nitrogen source and fixer in organic
agriculture [4].
Gliricidia plant extracts showed high potent anthelmintic effects in the studies per-
formed by Romero et al. [47]. The active compounds in gliricidia comprise glycosylated
flavonoids, methoxy phenols, phenylpropanoids, anthraquinone glycosides, amino acids,
and glycosylated phenolic acids. These compounds represent an alternative method to
control gyrodactylid infections in tilapia [48].
The particle size distribution was 79% less than 500 µm and 21% greater than 500 µm
for cassava leaf flour (Figure 5(A1)), 95% and 5% for gliricidia leaf flour (Figure 5(B1)), and
90% and 10% for rice husk flour (Figure 5(C1)).
Figure 5(A2,A3) (cassava leaf flour), Figure 5(B2,B3) (gliricidia leaf flour) and Fig-
ure 5(C2,C3) (rice husk flour) show agglomerations between the particles below 500 µm,
while around the particles above 500 µm small particles are observed adhering. The above
effect can be associated with the drying process of the raw materials, the electrostatic
effects, and van der Waals forces. Agglomeration characteristics are widely desired for
extruded food processes in both human and animal feed [49]. An important application of
the agglomeration process is in the production of instant products in which the primary
particles are agglomerated to give a product in the form of a granule. In the production
of fish feed, agglomeration improves the characteristics of the extrudate in terms of its
Sustainability 2022, 14, 6059 9 of 17

Sustainability 2022, 14, x FOR PEER REVIEW 9 of 17

wettability, dispersibility, and dissolution [50]. Figure 6 shows the cohesion index of the
raw materials.
Figure
Figure 55shows
showsthe
the analysis
analysis of the
of the flourflour samples
samples analyzed
analyzed with
with the the stereomicroscope
stereomicroscope
showed variable particle sizes where most of them were below 0.5 mm
showed variable particle sizes where most of them were below 0.5 mm (500 µm). (500 µm).

Figure 5. Analysis of the flour samples analyzed with the stereomicroscope. (A1–A3) correspond to
cassava leaf flour; (B1–B3) to gliricidia leaf flour; and (C1–C3) to rice husk flour.

The particle size distribution was 79% less than 500 µm and 21% greater than 500 µm
for cassava leaf flour (Figure 5A1), 95% and 5% for gliricidia leaf flour (Figure 5B1), and
90% and 10% for rice husk flour (Figure 5C1).
Figure 5A2,A3 (cassava leaf flour), Figure 5B2,B3 (gliricidia leaf flour) and Figure
5C2,C3 (rice husk flour) show agglomerations between the particles below 500 µm, while
around the particles above 500 µm small particles are observed adhering. The above effect
can be associated with the drying process of the raw materials, the electrostatic effects,
and van der Waals forces. Agglomeration characteristics are widely desired for extruded
food processes in both human and animal feed [49]. An important application of the ag-
glomeration process is in the production of instant products in which the primary parti-
cles are agglomerated to give a product in the form of a granule. In the production of fish
feed, agglomeration
Figure 5. Analysis of theimproves the characteristics
flour samples analyzed with theof stereomicroscope.
the extrudate in terms ofcorrespond
(A1–A3) its wettabil-
to
Figure 5. Analysis and
ity, dispersibility, of the flour samples analyzed
dissolution with the stereomicroscope.
index of(A1–A3)
the rawcorrespond to
cassava leaf flour; (B1–B3) to gliricidia[50]. Figure
leaf flour; 6 shows
and (C1–C3)the cohesion
to rice husk flour. ma-
cassava
terials. leaf flour; (B1–B3) to gliricidia leaf flour; and (C1–C3) to rice husk flour.
The particle size distribution was 79% less than 500 µm and 21% greater than 500 µm
for cassava leaf flour (Figure 5A1), 95% and 5% for gliricidia leaf flour (Figure 5B1), and
90% and 10% for rice husk flour (Figure 5C1).
Figure 5A2,A3 (cassava leaf flour), Figure 5B2,B3 (gliricidia leaf flour) and Figure
5C2,C3 (rice husk flour) show agglomerations between the particles below 500 µm, while
around the particles above 500 µm small particles are observed adhering. The above effect
can be associated with the drying process of the raw materials, the electrostatic effects,
and van der Waals forces. Agglomeration characteristics are widely desired for extruded
food processes in both human and animal feed [49]. An important application of the ag-
glomeration process is in the production of instant products in which the primary parti-
cles are agglomerated to give a product in the form of a granule. In the production of fish
feed, agglomeration improves the characteristics of the extrudate in terms of its wettabil-
ity, dispersibility, and dissolution [50]. Figure 6 shows the cohesion index of the raw ma-
Figure 6.
Figure 6. Cohesion
Cohesionindexindexof of
raw materials.
raw materials.
terials.
The cohesion
The cohesionindex
indexforfor
cassava leafleaf
cassava flour, gliricidia
flour, leaf flour,
gliricidia leaf and rice
flour, husk
and flour
rice were
husk flour were
19.22, 24.14, and 20.29, respectively. According to IC-based powder classification
19.22, 24.14, and 20.29, respectively. According to IC-based powder classification [24], all [24], all
flour samples in this study are classified as extremely cohesive powders. The cohesion
flour samples in this study are classified as extremely cohesive powders. The cohesion
index obtained is consistent with the images obtained using the stereomicroscope where
index obtained is consistent with the images obtained using the stereomicroscope where
they show a high attraction between particles. Similarly, cohesion indices greater than 19
they show a high attraction between particles. Similarly, cohesion indices greater than 19
are evidence of large inter particulate cohesion forces such as solid bridges, liquid bridges,
van der Waals interactions [51], electrostatic forces and mechanical interlocks [49].
The cohesion index results confirm that the flours can be used in the elaboration of
animal feed, especially in extruded foods, since a reduction in the particle size also increases
the contact area between the particles, thus increasing the cohesive forces [24].
The results of particle size and cohesion indexes show that the flours obtained to
Figure 6. with
comply Cohesion
theindex of raw materials.
necessary parameters for the elaboration of extruded food for fish [50].
Figure 7 shows the results of the analysis of powders at different speeds.
The cohesion index for cassava leaf flour, gliricidia leaf flour, and rice husk flour were
The forces involved in the powder flow are gravity, friction, cohesion (attraction
19.22, 24.14, and 20.29, respectively. According to IC-based powder classification [24], all
between particles), and adhesion (attraction between the particle and the walls of the
flour samples in this study are classified as extremely cohesive powders. The cohesion
container). The flow curve of each sample was obtained during the compression and
index obtained is consistent with the images obtained using the stereomicroscope where
decompression
they show a hightest by performing
attraction three cycles
between particles. for each
Similarly, powder
cohesion sample,
indices greaterwithout
than 19 adding
any binder liquid.
Figure 7 shows that all samples follow a similar pattern on the graphs of the three
compression and decompression cycles indicating reproducible behavior for each powder
 Sustainability 2022, 14, x FOR PEER REVIEW 10 of 17

Sustainability 2022, 14, 6059 10 of 17

are evidence of large inter particulate cohesion forces such as solid bridges, liquid bridges,
van der Waals interactions [51], electrostatic forces and mechanical interlocks [49].
duringThethe
cohesion
test. index results
The area ofconfirm that the flours
the compression can is
zone be always
used in the elaboration
higher of of the
than that
animal feed, especially in extruded foods, since a reduction in the particle size also
decompression zone for all samples, indicating that less energy is required to decompress in-
creases
the the contact
powder area between
compared the particles,This
to compression. thus increasing the cohesive
shows greater bondingforces [24]. hydrogen
forces,
The results of particle size and cohesion indexes show that the flours obtained to
bonds, and Van der Walls forces within the particle and fewer forces acting between the
comply with the necessary parameters for the elaboration of extruded food for fish [50].
particles [52].
Figure 7 shows the results of the analysis of powders at different speeds.

Figure 7.
Figure 7. Powder
Powderanalysis
analysisat at
different speeds.
different speeds.

The forces
The maximum involved
peakinforce
the powder
obtained flow arecassava
with gravity, leaf
friction,
flourcohesion
is around(attraction be-gliricidia
2.6 N, for
tween particles), and adhesion (attraction between the particle and the walls of the con-
leaf flour around 2 N and rice husk flour around 4.8 N. On the other hand, lower force
tainer). The flow curve of each sample was obtained during the compression and decom-
peaks indicate that more energy is required for the compression of rice husk flour than for
pression test by performing three cycles for each powder sample, without adding any
cassava leaf flours and gliricidia leaf flour, indicating greater stiffness between the flour
binder liquid.
particles
Figureof rice huskthat
7 shows compared
all samples to other
followflours.
a similar pattern on the graphs of the three
Gliricidia leaf flour, having lower
compression and decompression cycles indicating force peaks, evidences
reproducible the presence
behavior for eachofpowder
protein and its
ease
during the test. The area of the compression zone is always higher than that of the decom-moisture,
of water absorption, acting as hydrocolloids and, therefore, quickly absorbing
this is consistent
pression zone for allwith the protein
samples, results
indicating thatobtained
less energy (25.63%) [53].
is required to The high force
decompress the peak in
rice huskcompared
powder flour (4.8toN)compression.
indicates the Thispresence of highbonding
shows greater amounts of cellulosic
forces, hydrogen compounds
bonds, that
and Van der Walls forces within the
provide greater rigidity to the macroparticles. particle and fewer forces acting between the particles
[52]. Powder analysis results confirm that the particle size of the raw materials can play
The maximum
an important role inpeak forceofobtained
terms with cassava
the parameters leaf extrusion
of the flour is around
system2.6 N,
andforthe
gliri-
quality of
cidia leaf flour around 2 N and rice husk flour around 4.8 N. On the
the granules. The correspondence between particle size, cohesion index, extrusion system other hand, lower
force peaks indicate that more energy is required for the compression of rice husk flour
parameters, and extrudate quality have already been reported for other extruded foods
than for cassava leaf flours and gliricidia leaf flour, indicating greater stiffness between
and food products [54]. The results obtained in the analysis of the powder for the flours,
the flour particles of rice husk compared to other flours.
as well as theleaf
Gliricidia results
flour, of the previous
having lower forceanalysis, indicatethethat
peaks, evidences it is possible
presence of proteintoandelaborate
extruded
its ease offish feeds
water with adequate
absorption, acting asphysical characteristics
hydrocolloids in terms
and, therefore, of sinking,
quickly floating and
absorbing
degradation
moisture, this[50].
is consistent with the protein results obtained (25.63%) [53]. The high force
peak in rice husk flour (4.8 N) indicates the presence of high amounts of cellulosic com-
3.2. Development
pounds that provide of Fish Food
greater rigidity to the macroparticles.
An affordable quality feed will make fish farming attractive to private investors and
increase fish production. When evaluating ingredients for use in the aquaculture sector,
several aspects deserve attention such as ingredient availability, accessibility, and nutrient
composition. The basic nutrient that cannot be compromised in the choice of ingredients
for the formulation and production of food is a protein [15].
In this research, three formulations were developed that provide nutritional require-
ments according to the availability of the raw material at each time of the year, regardless
of the abundance or scarcity of it, so that fish feed can be produced at any time of the year.
 An affordable quality feed will make fish farming attractive to private investors and
increase fish production. When evaluating ingredients for use in the aquaculture sector,
several aspects deserve attention such as ingredient availability, accessibility, and nutrient
composition. The basic nutrient that cannot be compromised in the choice of ingredients
for the formulation and production of food is a protein [15].
Sustainability 2022, 14, 6059 11 of 17
In this research, three formulations were developed that provide nutritional require-
ments according to the availability of the raw material at each time of the year, regardless
of the abundance or scarcity of it, so that fish feed can be produced at any time of the year.
The
Thefish
fishfeed
feedformulation
formulationshownshownin inTable
Table22was wasmade
madebased
basedononthethenutritional
nutritionalneeds
needs
of Red Tilapia, although it can be used for other fish
of Red Tilapia, although it can be used for other fish species. species.
Formulation
Formulation33was wastaken
takenforforthe
theanalyses,
analyses,which
whichprovides
providesthethenecessary
necessarynutrients
nutrientsforfor
fattening Tilapia: Spirulina sp. (20%), cassava leaf flour (50%), gliricidia leaf
fattening Tilapia: Spirulina sp. (20%), cassava leaf flour (50%), gliricidia leaf flour flour (20%)(20%)
and
flour of rice
and flour ofhusk (10%).
rice husk (10%).
The
The pellets weremade
pellets were madefollowing
followingthe theformulation.
formulation. The
The procedure
procedure allowed
allowedthethe
incorpo-
incor-
ration of air into the pellets and the obtaining of semi-smooth walls that
poration of air into the pellets and the obtaining of semi-smooth walls that trapped airtrapped air inside
the pellet,
inside thethus facilitating
pellet, the floating
thus facilitating process,process,
the floating as shown as in Figure
shown in 8.
Figure 8.

Figure8.8.Fish
Figure Fishfood
foodaspect
aspectand
andbehavior.
behavior.(A):
(A):Pellets,
Pellets,(B):
(B):Floating,
Floating,(C):
(C):Stability.
Stability.

Tomeasure
To measurethe thefloating
floatingof ofthe
thepellet,
pellet,aameasurement
measurementwas wastaken
takenof ofthe
thetime
timein inminutes
minutes
it takes
it takes for the
the pellet to sink, in this sense, 30% of the pellets sank during the firsts,10
pellet to sink, in this sense, 30% of the pellets sank during the first 10 10% s,
floated
10% for 20
floated formin andand
20 min the the
remaining
remaining 60%60%of the pellets
of the pelletsfloated
floated forfor
more
more than
than4040
min.
min. It
Itwas
wasobserved
observed that
thatthe moisture
the moisture of of
thethe
pellet during
pellet duringextrusion
extrusion andandthethefinal drying
final of the
drying of
pellet
the influence
pellet influence on the
on thefloating
floatingtimes, thethe
times, combination
combination between
between extrusion
extrusion temperature
temperature be-
tween 110
between 110°C◦ Cand
and120120°C ◦ Cand
andthe thefinal
finaldrying
dryingof ofthe
the pellets
pellets at 60 °C ◦ C until achieving
achieving aa final
final
moisture
moisturebetween
between10–15%10–15% in in dry
dry weight,
weight, generate
generate long
long floating
floating times,
times, onon thethe other
other hand,
hand,
low
lowtemperatures
temperaturesduring duringthe theextrusion
extrusionprocess
processprevent
preventthethegeneration
generationof ofwater
watervapor
vaporthat
that
generates
generatesporosities
porositiesinside
insidethe thepellet,
pellet,porosities
porositieswhere
wheresmall
smalltraps
trapsarearetrapped
trappedair airparticles
particles
that
thatfavors
favorsthe thefloating
floatingofofthe themixture
mixturegenerated
generated ananincreasing
increasing trend
trendin the
in thefloating time
floating of
time
the pellets
of the [38].
pellets Fish
[38]. feed
Fish andand
feed its tests areare
its tests shown
shownin Figure
in Figure8. 8.
Comparing
Comparing the the results
results of of the
the feed
feed for
forfish
fishin
inpellets
pelletsdeveloped
developedin inthis
thisresearch
researchwith
with
the
the nutritional contribution of the formulation with the nutritional requirements of
nutritional contribution of the formulation with the nutritional requirements of the
the
tilapia in the growth stage are widely covered both at the level of proximal composition, as
well as vitamins, amino acids, and minerals. The studies show that the Spirulina platensis
protein concentrate exhibits a high protein digestibility rate of 87.45–97.81% [18], as shown
un Table 4.
Table 4. Tilapia: Nutritional need vs supply pellets.

Proximal Composition In This Study In the Literature

Moisture 10–15% 21–24% [55,56]
Ashes 13% 11% [57–61]
Protein 0.035 kg 0.035 kg [59–62]
Fat 16% 10–15% [59,62,63]

The use of Spirulina sp. incorporates essential amino acids, proteins, polysaccharides.
The use of cassava leaf flour and gliricidia leaf flour are a good source of protein as
well as some amino acids, including methionine, arginine, isoleucine, and leucine. The use
of rice husk leaf flour incorporates, in addition to amino acids, the B vitamins and minerals,
which cover the nutritional needs of Tilapia [63].
Sustainability 2022, 14, 6059 12 of 17

Developed fish feed is a source of micronutrients such as vitamins, minerals, and

lipids. In addition, it contributes to improving the digestibility and palatability of any food,
which is particularly important in early-stage diets for many species [16].
The focus of fish production is to increase its unsaturated fatty acid content without
relying heavily on fish oils. Microalgae-based fish food is gaining attention and will become
a major industry in the coming years. Many small fish are being farmed with microalgae as
live prey or by aquaculture in green water in hatcheries [18].
Even though, there are currently various sources of animal feed such as use of pelagic
fish [64], soy-based feed [65], use of insects as feed [66], these animal feed sources are not
widely produced in Colombia. Therefore, the present concentrate becomes an ideal animal
feed source thanks to the fact that it takes advantage of the agro-industrial residues typical
of the fish farming regions of our country.

3.3. Pilot Production of Spirulina sp. to Guarantee the Sustainability of the Aquaculture Sector
A potential benefit of this research sought to reduce the total cost of fish production
in rural areas by reducing the cost of concentrate for fish incorporating agro-industrial
residues from the region, and adopt bioprocesses easy to operate by the community to
produce a local fish food enhanced with Spirulina sp.
The municipality of San Carlos, Antioquia, and specifically the area of El Jordán was
selected as a target location for the implementation of the results coming from
Sustainability 2022, 14, x FOR PEER REVIEW 13 this
of 17research,
as shown in Figure 9.

Figure 9.
Figure 9. Location
LocationofofElEl
Jordán, SanSan
Jordán, Carlos (Antioquia).
Carlos (Antioquia).

San Carlos
San Carlosisis
a municipality withwith
a municipality a fish afarming traditiontradition
fish farming that strategically has capac- has ca-
that strategically
ities for fingerling production. Specifically, the El Jordán area has an
pacities for fingerling production. Specifically, the El Jordán area has an averageaverage climate be- climate
tween 18–25 °C, ◦and the availability of water through its watersheds makes it optimal for
between 18–25 C, and the availability of water through its watersheds makes it optimal
fish production and establishing production chains.
for fish production and establishing production chains.
In contrast, this territory was affected by the armed conflict during the 1990s, which
In contrast,
generated povertythis
andterritory was affected
backwardness byand
in its social the economic
armed conflict duringCurrently,
development. the 1990s, which
generated
the region is experiencing post-conflict economic growth, where technology adoption Currently,
poverty and backwardness in its social and economic development. is
the region
required is experiencing
to strengthen post-conflict
value chains such as fisheconomic
farming growth,
and therebywhere technology
generate job oppor-adoption
istunities
required to rural
for the strengthen value chains such as fish farming and thereby generate job
community.
opportunities for the
Two pilot were rural community.
formulated for its implementation by the rural community of the mu-
nicipality of San Carlos to guarantee
Two pilot were formulated for the its
sustainability of this research:
implementation by the rural community of the
•
municipality
Productionof of
San Carlossp.
Spirulina to as
guarantee the by
raw material sustainability
the farmers of this research:
•• Production ofoffish
Production food incorporating
Spirulina the Spirulina
sp. as raw material produced.
by the farmers
• A model of social appropriation of knowledge was
Production of fish food incorporating the Spirulina implemented
produced. in which the re-
search group CIBIOT of the Universidad Pontificia Bolivariana installed a set of five bio-
A model of social appropriation of knowledge was implemented in which the research
reactors equal to the number of families in rural areas of the municipality of San Carlos
group CIBIOT of the Universidad Pontificia Bolivariana installed a set of five bioreactors
and trained them in the methodology to produce microalgae as discussed in this article.
Each bioreactor was 315 L capacity, coupled with a solar panel as renewable energy sys-
tem for the operation of an air pump that supplies aeration to the system, as shown in
Figure 10.
 tunities for the rural community.
Two pilot were formulated for its implementation by the rural community of the mu-
nicipality of San Carlos to guarantee the sustainability of this research:
• Production of Spirulina sp. as raw material by the farmers
Sustainability 2022, 14, 6059 • Production of fish food incorporating the Spirulina produced. 13 of 17
A model of social appropriation of knowledge was implemented in which the re-
search group CIBIOT of the Universidad Pontificia Bolivariana installed a set of five bio-
reactors equal to the number of families in rural areas of the municipality of San Carlos
equal to the them
and trained number of methodology
in the families in rural areas ofmicroalgae
to produce the municipality of San
as discussed Carlos
in this and trained
article.
them in the methodology
Each bioreactor to produce
was 315 L capacity, microalgae
coupled as discussed
with a solar panel as in this article.
renewable Each
energy bioreactor
sys-
was
tem 315 L capacity,
for the coupled
operation with
of an air a solar
pump that panel as renewable
supplies aeration to energy system
the system, for theinoperation
as shown
of an air
Figure 10.pump that supplies aeration to the system, as shown in Figure 10.

Figure 10. Schema for sustainable production of Spirulina sp. in rural areas. Inoculum preparation
and pilot production in 350 L bioreactor.

After generating the infrastructure conditions and the appropriation of knowledge to

produce Spirulina sp., a model of community production of fish food is proposed. Under
this model, each farmer produces Spirulina sp. on his farm, stores and transports it to a
collection center where the fish feed production process is carried out by processing the
agro-industrial residues of cassava leaf flours, gliricidia leaves, and rice husk, which are
present in the region.
Under this model, farmers recognize that a job opportunity is created that comple-
ments their free time and adds value to the family economy. With the development of this
pilot test, it was estimated that the time a farmer invests in the production of Spirulina sp.
is equivalent to two days of work each month. The model is schematized in Figure 11.

Figure 11. Model for fish food production from agro-industrial residues enhanced with locally
produced Spirulina sp. in the rural community of El Jordán, Antioquia.
Sustainability 2022, 14, 6059 14 of 17

4. Conclusions
The composition of the cultivated Spirulina sp. presented a percentage of 6.79% ± 0.05
of moisture, 6.93% ± 0.01 of ash, 66.88% ± 0.33 of protein and 5.50% ± 0.26 of fat, which
allows it to be used as fish food, especially Tilapia since they require a source of protein of
35–45% and fat of 10–15%.
The results of the characterization of cassava leaf flours, gliricidia leaves, and rice husk
show that this mixture of these three raw materials plus spirulina, has great application
in the production of pellet-type fish food, due to its proximal composition are within the
reference values, more than 79% of the particles are less than 500 µm, the cohesion index
of for cassava leaf flour, gliricidia leaf flour, and rice husk flour was 19.22, 24.14 and 20.29
respectively and the results of the analysis of powders confirm that the particle size of the
raw materials can play an important role in terms of the parameters of the extrusion system
and the quality of the granules.
It was possible to produce fish food in the form of pellets using Spirulina sp. and agro-
industrial waste flours: cassava leaves, gliricidia leaves and rice husk with 60% floating for
more than 40 min, 80% of the pellets kept their shape for 4 h, keeping the micronutrients
longer in the pellets.
Two pilot were formulated for its implementation by the rural community of the
mu-nicipality of San Carlos to guarantee the sustainability of this research, comprising the
production of Spirulina sp. as raw material by the farmers and therefore the production of
fish food incorporating the Spirulina sp. produced. The model proposed opportunities for
economic growth and the implementation of green technologies for rural families.

Author Contributions: Conceptualization, M.R.-C., D.S.-O., C.O.-L. and L.R.-C.; Methodology,

M.R.-C., C.O.-L., D.S.-O. and L.R.-C.; validation, M.R.-C., L.R.-C., C.O.-L. and D.S.-O.; formal analysis,
M.R.-C., L.R.-C., D.S.-O. and C.O.-L.; investigation, M.R.-C., D.S.-O., C.O-L. and L.R.-C.; resources,
M.R.-C.; data curation, C.O.-L.; writing—original draft preparation, M.R.-C., D.S.-O., C.O-L. and
L.R.-C.; writing—review and editing, M.R.-C., L.R.-C. and C.O.-L.; visualization C.O.-L.; funding
acquisition, M.R.-C. All authors have read and agreed to the published version of the manuscript.
Funding: This research was funded by MINCIENCIAS and The Government of Antioquia, grant
number 80740-009-2019 and the APC was funded by Universidad Pontificia Bolivariana.
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: Not applicable.
Conflicts of Interest: The authors declare no conflict of interest.

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