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© 2024 by the authors; licensee Asian Online Journal Publishing Group 
 

Agriculture and Food Sciences Research 
Vol. 11, No. 2, 136-145, 2024 

ISSN(E) 2411-6653/ ISSN(P) 2518-0193 
DOI: 10.20448/aesr.v11i2.6227 

© 2024 by the authors; licensee Asian Online Journal Publishing Group 

 
 

 
 
 
Encapsulation of vegetable oils in polylactic acid nanofibers to improve oil 
retencion in feed aquaculture 

 
Aline Moreira de Alcantara1   

Tulio Pacheco Boaventura2   

Renan Rosa Paulino3   

Priscila Vieira e Rosa4   

Julio Cesar Ugucioni5   

Juliano Elvis Oliveira6   

 

 
( Corresponding Author) 

 
1,2Federal University of Lavras, Lavras, Minas Gerais, Brazil. 
1Email: Alcântara@ufla.com.br  
2Email: tuliopb1@hotmail.com  
3,4Department of Animal Science, Federal University of Lavras, Lavras, Minas Gerais, Brazil. 
3Email: renan.paulino@ufla.br  
4Email: priscila@ufla.br  
5Department of Physics, Federal University of Lavras, Lavras, Minas Gerais, Brazil. 
5Email: Julio.Ugucioni@ufla.br  
6Department of Engineering, Federal University of Lavras, Lavras, Minas Gerais, Brazil. 
6Email: juliano.oliveira@ufla.br  

 
Abstract 

The feed used in fish farming contains lipids that erode and disperse in water, adversely affecting 
fish nutrition and contributing to water pollution. This study evaluated the encapsulation of corn 
and gold linseed oils in polylactic acid (PLA) nanofibers produced via Solution Blow Spinning. 
The nanofibers were characterized using Fourier-Transform Infrared (FTIR) spectroscopy, 
Scanning Electron Microscopy (SEM), contact angle analysis, and encapsulation efficiency tests. 
Additionally, the release profiles of the oils were studied under simulated tank water and 
gastrointestinal conditions. The results demonstrated successful encapsulation of the oils within 
the PLA matrix. FTIR analysis confirmed the presence of oil in the nanofibers (wavenumbers 
2925 and 2853 cm-1), with encapsulation efficiencies ranging from 81.54±10.36% to 
99.14±4.55%. SEM revealed uniform nanofiber morphology with smooth surfaces and average 
diameters between 157±49 and 385±133 nm. All nanofibers exhibited contact angles above 90°, 
indicating hydrophobic behavior. Feed samples containing free oils showed significantly higher 
lipid release into water compared to feed incorporating encapsulated oils. The findings suggest 
that encapsulating vegetable oils in PLA nanofibers reduces lipid dispersion in water while 
maintaining nutrient availability. This polymeric system offers a sustainable alternative to 
enhance aquaculture feed efficiency and reduce environmental impacts, contributing to cleaner 
water and healthier fish. 
 

Keywords: Farming, Formulation, Nutricion, Oil leakage rate, Pellet, Solution blow, Spinning. 

 
Citation: |Alcantara, A. M. de, Boaventura, T. P., Paulino, R. R., 
Rosa, P. V. e, Ugucioni, J. C., & Oliveira, J. E.  (2024). Encapsulation 
of vegetable oils in polylactic acid nanofibers to improve oil 
retencion in feed aquaculture. Agriculture and Food Sciences 
Research, 11(2), 136–145. 10.20448/aesr.v11i2.6227  
History:  
Received: 31 October 2024 
Revised: 6 December 2024 
Accepted: 10 December 2024 
Published: 20 December 2024 
Licensed: This work is licensed under a Creative Commons 

Attribution 4.0 License  
Publisher:  Asian Online Journal Publishing Group 
 

Funding: This research is supported by the Conselho Nacional de 
Desenvolvimento Científico e Tecnológico-CNPq Brazil (Grant number: 
CNPq-Brasil –150480/2023-7) and Fundação de Amparo a Pesquisa de Minas 
Gerais–FAPEMIG (Grant number: FAPEMIG-Brasil –APQ-05593-24, 
BPD-00406-22, APQ-02146-21), and Coordenação de Aperfeiçoamento de 
Pessoal de Nível Superior – CAPES-Brazil. 
Institutional Review Board Statement: Not applicable. 
Transparency: The authors confirm that the manuscript is an honest, 
accurate, and transparent account of the study; that no vital features of the 
study have been omitted; and that any discrepancies from the study as 
planned have been explained. This study followed all ethical practices during 
writing. 
Competing Interests: The authors declare that they have no competing 
interests. 
Authors’ Contributions: Conceptualization, methodology, validation, formal 
analysis, investigation and writing – original draft, A.M.d.A., T.P.B. and 
R.R.P.; methodology, validation, formal analysis, investigation, resources, 
data curation, investigation and writing, P.V.e.R. and J.C.U.; methodology, 
validation, formal analysis, investigation, resources, data curation, 
investigation, reviewing and editing, visualization, supervision, project 
administration and funding acquisition, J.E.d.O. All authors have read and 
agreed to the published version of the manuscript. 

mailto:Alcântara@ufla.com.br
mailto:tuliopb1@hotmail.com
mailto:renan.paulino@ufla.br
mailto:priscila@ufla.br
mailto:Julio.Ugucioni@ufla.br
mailto:juliano.oliveira@ufla.br
https://creativecommons.org/licenses/by/4.0/
https://creativecommons.org/licenses/by/4.0/
https://www.doi.org/10.20448/aesr.v11i2.6227
https://orcid.org/0000-0002-7343-2280
https://orcid.org/0000-0002-6143-5417
https://orcid.org/0000-0001-7915-5694
https://orcid.org/0000-0002-0052-079X
https://orcid.org/0000-0002-6362-5919
https://orcid.org/0000-0001-9570-8308


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Contents 
1. Introduction .................................................................................................................................................................................... 137 
2. Materials and Methods ................................................................................................................................................................. 138 
3. Results .............................................................................................................................................................................................. 139 
4 Discussions ....................................................................................................................................................................................... 142 
5. Conclusion ....................................................................................................................................................................................... 143 
References ............................................................................................................................................................................................ 144 
 

 

Contribution of this paper to the literature 
This article proposes the use of nanotechnology as a sustainable solution for incorporating 
vegetable oil into fish feed, enhancing fish nutrition while reducing nutrient waste and water 
contamination. 

 

1. Introduction 
The fish feed should contain proteins, lipids, vitamins, and carbohydrates for efficient maintenance of adequate 

animal growth within the shortest time [1]. It is important to give special attention to the balance between intake, 
retention, and excretion, because excessive intake of certain nutrients is responsible for intoxication events and, 
consequently, water pollution [2, 3]. Water contamination from fish farms can occur due to therapeutic residues, 

fish pathogens, and feed‐derived wastes, and the loss of these ingredients is also related to economic disadvantages 
[4]. Depending on production systems in fish farming and particularly the kind of effluent treatment, the waste 
load per ton of fish production can become a serious environmental challenge [3]. Monitoring and treating this 
feed waste in effluent directly is an inaccurate and costly process. The remainder of the undigested feed is dispersed 
on water (fat, vitamins, carbohydrates, and proteins) or excreted in the feces as solid waste, and the byproducts of 
fish metabolism are excreted as dissolved waste (ammonia, urea, phosphate). Concerns about the oils used in feed 
fortification also include how their waste will affect water quality since they are highly related to aquatic pollution. 
Oil leaching can lead to the formation of a mechanical barrier in the water surface, preventing light and oxygen 
from penetrates the water column and consequently reducing photosynthesis and phytoplankton formation. The 
decrease of phytoplankton in the aquatic medium is also responsible the reducing oxygen supply to the water 
column [5]. Therefore, the formulation of diets with low environmental impact is necessary to avoid damage to 
water quality in aquaculture production systems [6]. In this context, the use of controlled release systems 
containing natural products in feed formulation is an alternative to reduce water contamination from fish farming.  

Fish meal and oil play an important role as lipid source in the fish diet. Because of their rich content in highly 
unsaturated fatty acids, particularly eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids, they act as 
nutritional supplements [2]. Essential unsaturated fatty acids promote fish growth by regulating metabolism and 
improving immunity. The problem is that the overuse of fish meal and oil in farmed fish diets results in higher 
nitrogen and phosphorus discharges into the water systems, which causes eutrophication [7, 8]. Moreover, these 
products are very expensive and obtained from finite sources, therefore their use represents not only an 
environmental but also an economic problem. Focused on developing more sustainable aquaculture processes 
several authors have been studying alternative lipid sources to replace fish oil [9, 10]. Gold linseed oil (LO) and 
corn oil (CO) represent a good alternative to fulfill this role, since they are a sustainable source of fatty acids that 
are metabolized by the fish organism, leading to the formation of long-chain polyunsaturated essential fatty acids of 
the n-3 (DHA and EPA) and n-6 (arachidonic acid) series, respectively. Also, both corn and gold linseed oils are 
highly digested by all fish species. Another advantage of their use is that they do not contain dioxins and pollutants 
[11] and some studies have demonstrated that these oils would improve the diet conversion rate of fish as well as 
fish health [12]. In recent years, there has been an increasing number of authors in the literature that studying the 
possibility of improving the composition and quality of animal feed using nanostructured materials in feed 
formulations [13-17]. Oral administration of nutrients to farmed fish brings inherent challenges that must be 
considered when designing a nanostructured carrier. Furthermore, each compartment of the gastrointestinal tract 
has a unique environment that includes its complement of enzymes and specific pH levels. Nanofibers can overcome 
these challenges and deliver their nutrients to the intestine, where most of the absorption occurs, then traveling to 
the bloodstream and finally being distributed through all the organism [18]. 

Techniques for obtaining polymeric nanofibers have been developed in the last decade. Solution blow spinning 
(SBS) is a fiber manufacturing process that uses two parallel concentric nozzles. Nanofiber production by SBS 
depends on several factors such as concentration and viscosity of the polymeric solution, ejection rate, air pressure, 
and working distance. These variables have a direct influence on the fiber production rate and fiber morphology 
[19-21]. In comparison with other techniques, such as electrospinning, the SBS technique has a higher ejection 
rate, fiber production, and versatility regarding the use of solvents, polymers, and applications, in addition to its 
safety and low cost [22, 23]. Nanofibers obtained by the SBS technique have several applications in animal science 
like as controlled delivery of progesterone to control the estrus cycle in livestock animals [24] and encapsulation 
of diclofenac sodium to create in situ devices for pododermatitis treatment [25]. 

Thus, it is notable that immediate improvements in fish feed formulations are needed for a more efficient 
administration of nutrients, which involves reducing waste and water contamination from fish farming and 
increasing the biodisponibility of nutrients in the intestinal tract. With this aim, we now report the development 
and physicochemical characterization of hydrophobic polylactic acid (PLA) nanofibers containing different ratios of 
gold linseed and corn oil, produced by the Solution Blow Spinning Technique. We posit that biodegradable and 
hydrophobic nanofibers could protect fatty acid against release in the aquatic environment and stomach chamber, 
improving fatty acid release effectiveness and, thus, creating the potential for a smart fish feed with the lowest 
environmental impact. 

 



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2. Materials and Methods 
2.1. Materials 

Polylactic acid (3251 D, 66KDa, Chemical Abstracts Service number 9051-89-2) was obtained from Nature 
Works (USA). Corn oil (brand Lizza), gold linseed oil (brand Pazze), and the materials used in feed production 
were purchased locally. Chloroform (Chemical Abstracts Service number 67-66-3) was obtained from Synth 
(Brazil).  The lipophilic dye Sudan III (Chemical Abstracts Service number 85-86-9) was purchased from Êxodo 
Científica (Brazil). 

 

2.2 Solution and Fiber Production 
The polylactic acid solution was prepared as follows: 12% (w/v) PLA was dissolved in chloroform under 

continuous stirring for 40 minutes. After the complete polymer dissolved, 50% (w/w) of vegetable oils were added 
to the solution [26, 27]. All the different fractions of corn oil (CO) and gold linseed oil (LO) used are shown in 
Table 1. The solution was kept under stirring and at room temperature for another 15 minutes. Finally, 0.125% 
(w/w) of the lipophilic dye Sudan III was added, and it were kept under stirring for 15 minutes. 
 
Table 1. Different fractions of corn oil and gold linseed oil used to formulate the polylactic acid solutions that will pass through the solution 
blow spinning process. 

Sample code Corn oil (%v/v) Linseed oil (%v/v) 

CO:LO – 100:0 100 0 
CO:LO – 75:25 75 25 
CO:LO – 50:50 50 50 
CO:LO – 25:75 25 75 
CO:LO – 0:100 0 100 
PLA nanofibers 0 0 

 
The fibers were obtained using the Solution Blow Spinning technique. The spinning system was formed 

consisted of a source of compressed air (CHIAPERINI, model MC 12 BPV,150 liters), a polymer injection pump 
(NEW ERA PUMP SYSTEMS, model Syringe Pump AL1000), and a glass syringe (20 mL and 19.33 mm). A 
rotating horizontal cylinder covered with aluminum paper was used to collect the nanofibers. The experimental 
apparatus was isolated inside a glass front-opening wooden box to maintain moisture (40-50%) and temperature 
control (approximately 30°C, using a reflective incandescent light bulb – TOVALIGHT E.27) throughout the 
process.  

The process took 3 hours for each sample and the spinning parameters used were adjusted based on the 
methodologies Nepomuceno, et al. [27] and Bonan, et al. [26]: feed rate – 6.00 mL/h; working distance – 20 cm; 
protrusion between the two concentric nozzles – 0.4 mm; air pressure – 172.37 kPa. The fibers were collected and 
stored for subsequent characterization. 

 

2.3. Fiber Characterization 
The absorption spectrum of the samples was obtained by Fourier-Transform Infrared Spectroscopy (FTIR) 

with the Attenuated Total Reflection (ATR) sampling technique. A total of 32 scans were considered for a 
wavenumber range of 4000-400 cm-1 with a resolution of 4 cm-1  [28]. 

Nanofibers were coated with a gold layer by sputtering (Balzers, Sputter Coating Device 050) for the analysis 
of their morphology using a scanning electron microscope (SEM). The SEM system used was a LEO EVO 40 
model (Carl Zeiss) equipped with Extended Variable Pressure (XVP), Bruker’s Quantax EDS for X-ray 

microanalysis, and a cryosystem from Gatan. For each experiment, the average fiber diameter was determined with 
the image analysis software Image J, using approximately 100 measurements randomly taken from the samples 
[29]. 

The hydrophobicity/hydrophilicity of the nanofibers was evaluated through contact angle measurement 
immediately, 5 seconds, and 60 seconds after the injection of a sessile drop on the nanofiber. The experiment was 
performed with a Kruss Drop Shape Analyzer Goniometer – DSA25 (Hamburg, Germany).  

To determine encapsulation efficiency, 30 mg of the nanofibers were dissolved in 10 mL of chloroform and the 
solution was quantified in a UV-Vis (Ultraviolet-Visible Spectroscopy) spectrophotometer (Shimadzu, model UV-
2601) at a wavelength of 513 nm. This process was performed in 5 different regions of each sample and the 
encapsulation efficiency was determined through Equation 1 (Designation). 

𝐸𝐸 (%) =  
𝐶𝐸

𝐶𝐼
⁄ 𝑥 100%                                           (1) 

Where: CE = Concentration of Sudan III encapsulated in nanofibers (Average calculated using the 5 regions); 
CI = Concentration of Sudan III initially added to the nanofibers. 

 

2.4. Feed Production 
A formulation simulating a conventional feed for omnivorous fish in the growth phase (the largest group of fish 

cultivated in the world) was produced: 45% soybean meal, 34.8% cornflour, 11.5% corn gluten meal 60, 5% corn 
oil/gold linseed oil, 1.5% dicalcium phosphate, 1.5% mineral and vitamin supplement, 0.5% sodium chloride, 0.19% 
DL-methionine, 0.125% Sudan III (a chemical marker used to track the release of vegetable oils), and water was 
added until the mixture obtain the proper consistency. The mixture was pelletized in an extruder (Adamo) and the 
product was dehydrated in a laboratory oven at 50°C for 24 hours [30].  Six feed samples were produced, 
containing contained the same corn oil/gold linseed oil fractions proposed in Table 1. 

 

2.5. Assessment of Nutrient Liberation in an Aqueous Medium (In Vitro) 
Approximately 30 mg of feed and feed containing nanofibers were placed into a falcon tube containing 3 mL of 

water, 1% (v/v) Tween 80, sodium bicarbonate (40 mg/L), and a pH equal to 7.0 [31]. The tubes were kept in an 

Note: Polylactic acid (PLA), Corn oil (CO) and Gold linseed oil (LO). 



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incubator under at 25°C and 120 rpm [32]. This process represents the preparation of one aliquot that was 
subsequently analyzed in a UV-Vis spectrophotometer (Shimadzu, model UV-2601) around at wavelength of 534 
nm. For each sample, 10 aliquots were prepared and collected after 10 minutes. 

 

2.6. Assessment of In Vitro Digestibility 
The Uv-Vis spectrophotometer (Shimadzu, model UV-2601) was used to measure absorbance around a 

wavelength of 534 nm. Tests of in vitro digestibility were based the methodology Wang, et al. [32]. The simulated 
gastric fluid of fish was prepared as follows: 20 mg of feed and feed containing nanofibers were added to an 
Erlenmeyer containing 7 mL of a sodium chloride (0.15 M) solution, 5% (w/v) of pepsin, and 1% (v/v) of Tween 
80; the solution was posteriorly acidified with HCl until the pH of 2.0. The solution was maintained in an incubator 
for 1 hour in the dark (the recipients were covered with aluminum paper to prevent the passage of ambient light), 
at 25°C, and 100 rpm. 

The simulated intestinal fluid of fish was prepared as follows: 20 mg of feed and feed containing nanofibers 
were added to an Erlenmeyer flask containing 7 mL of a sodium chloride (0,15 M) solution, 1.5% (w/v) of 
pancreatin, 0.5% (w/v) of amylase, 0.3% (w/v) of bile salts, and 1% (v/v) of Tween 80; the solution pH was 
posteriorly adjusted to 6.0. The solution was maintained in an incubator for 5 hours in the dark (the containers 
were covered with aluminum foil paper to prevent the passage of ambient light), at 25°C, and 40 rpm. 

The tests described in the previous paragraphs were also performed for the 6 feed samples containing different 
fractions of the free vegetable oils in its formulations. The presence of lipids in the solutions was evaluated using 
Uv-Vis spectrophotometer. 

 

2.7. Statistical Analysis 
All data were subjected to the Shapiro-Wilk normality test and Levene's test for homoscedasticity. Parametric 

data were analyzed by Variance Analysis (ANOVA) (One-way or Two- way) followed by Tukey's test at 5% 
probability. Past software were used for data analysis. 

 

3. Results 
Fiber characterization. 
 

3.1. Scanning Electron Microscopy (SEM) 
Figure 1 shows the SEM images obtained for all the nanofibers produced in this paper, demonstrating that 

they exhibited uniform morphology and smooth surface. Table 2 shows average diameter values for each sample, 
calculated using the software Image J.  
 
Table 2. Average fiber diameter and standard deviation of PLA nanofibers containing different fractions of vegetable oils. 

Sample CO: LO 100:0 CO: LO 75:25 CO: LO 
50:50 

CO: LO 
25:75 

CO: LO 
0:100 

PLA 
nanofibers 

Average diameter (nm) 168±52d 209±56c 385±133a 257±83b 157±49d 167±46d 
Note: The letters a, b, c, and d in the table indicate statistical differences between results. Results followed by different letters differ 

significantly between treatment (P<0.05). Ony-way ANOVA and Tukey’s test were used to determine statistical significance. 

 

 
Figure 1. Scanning electron microscopy of nanofibers containing: (a) CO: LO – 
100:0, (b) CO: LO - 75:25, (c) CO: LO - 50:50, (d) CO: LO - 25:75, (e) CO: LO – 
0:100, (f) PLA nanofibers. 

 
According to statistical analysis, higher values of average diameter were obtained for PLA nanofibers 

containing mixtures of corn oil and gold linseed oil (CL: LO – 50:50). The incorporation of pure oils did not affect 



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the samples average diameter and the minor values of average diameter were obtained for PLA nanofibers, CO: LO 
0:100 and CO: LO – 100:0. 

 

3.2 Fourier Transform Infrared Spectroscopy (FTIR) 
Figure 2 shows the FTIR spectra obtained from the different samples. The infrared analysis reveals peaks in 

the spectra of the various samples produced. Notable peaks were observed in the regions around 3000, 2925, and 
2853 cm-1. 

 

 
Figure 2. The infrared spectrum of PLA nanofibers containing corn oil and gold linseed oil in different proportions and of the 
pure vegetable oils in the wavenumber region from 4000 to 400 cm-1. This region corroborates the encapsulation of the 
vegetable oils. 

 

3.3. Contact Angle 
Table 3 shows the average contact angle values 60 seconds after the injection of water drop onto the surface of 

the nanofibers. All samples are hydrophobic since the contact angle was higher than 90° for all nanofibers. The 
highest contact angle were observed for the samples CO: LO – 50:150 and CO: LO – 25:75. The minors contact 
angle value were found for the samples PLA nanofibres and CO: LO – 0:100. 
 
Table 3. Contact angles of PLA nanofiber mat and PLA/Vegetable oils blend nanofiber mats with their standard deviations. 

Sample Average contact angle (°) 

CO: LO – 100:0 109.9±2.7 ac 

CO: LO – 75:25 116.7±7.4ab 

CO: LO – 50:50 106.3±1.8 c 

CO: LO – 25:75 107.8±3.4 c 

CO: LO – 0:100 114.5±2.7a 

PLA nanofibers 114.6±4.8a 

Note: The letters a, b and c in the table indicate statistical differences between results. Results followed by different letters differ significantly between 
treatment (P<0.05). Ony-way ANOVA and Tukey’s test were used to determine statistical significance. 

 

3.4. Encapsulation Efficiency 
Table 4 shows the encapsulation efficiency for each formulation. The results reveal that the CO: LO – 50:50 

sample had the lowest encapsulation efficiency (81.54±10.36%). The highest encapsulation efficiency was obtained 
for samples CO: LO –100:0 (99.14±4.55 %) and CO: LO – 0:100 (97.32±11.09 %). 

 
 
 
 
 



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Table 4. Encapsulation efficiency of polylactic acid nanofibers containing different fractions of vegetable oils. 

Sample Encapsulation efficiency (%) 

CO:LO – 100:0 99.14±4.55a 

CO:LO – 75:25 94.06±9.47abc 

CO:LO – 50:50 81.54±10.36b 

CO:LO – 25:75 85.97±4.72 abc 

CO:LO – 0:100 97.32±11.09 ac 

Note: The letters a, b and c in the table indicate statistical differences between results. Results followed by different letters differ significantly between 
treatment (P<0.05). Ony-way ANOVA and Tukey’s test were used to determine statistical significance. 

 

3.5. Assessment of Nutrient Liberation in an Aqueous Medium (In Vitro) 
In Figure 3 it is possible to observe the fraction of oil released into the water during 10 minutes of exposure of 

the different materials. When comparing the composition of the feed samples with and without oil encapsulated in 
nanofibers, it is observed that the CO: LO – 75:50 and CO: LO – 50:50 samples exhibited statistically similar oil 
release. The CO: LO – 100:0, CO: LO – 25:75, and CO: LO – 0:100 samples, containing oil encapsulated in 
nanofibers, showed lower oil release compared to the samples with free oil in the feed. When comparing oil release 
across different feed compositions, the CO: LO – 0:100 sample showed the highest release. For feeds with oil 
encapsulated in nanofibers, the CO: LO – 50:50 sample recorded the highest oil release. 

 

 
Figure 3. Comparison between the maximum quantity of vegetable oils released by the fish feed and feed containing 
nanofibers after 10 minutes in contact with an aqueous medium simulating fish tank condition. 
Note: Results followed by different capital letters differ significantly between the different treatment (Fish feed a fndeed containing 

nanofibers) of the same compositions (P<0.05).  Results followed by different lowercase letters differ significantly between the 
different compositions of the same treatment (P<0.05).  Two-way ANOVA and Tukey’s test were used to determine statistical 
significance. 

 
3.6. Assesment of in Vitro Digestibility 

Figure 4A shows the results of the digestibility test of the feeds in gastric fluid over a period of 1 hour. When 
comparing the same feed composition with and without oil encapsulated in nanofibers, it is observed that all feed 
samples without encapsulated oil exhibited higher digestibility compared to those containing encapsulated oil. 
When comparing the different feed compositions, the CO – 50:50 sample showed the highest digestibility. A similar 
pattern was observed for feeds with encapsulated oil, where the CO – 50:50 sample also presented the highest 
digestibility. 

Figure 4B presents the results of the digestibility test of the feeds in intestinal fluid over a period of 5 hours. 
When comparing the same feed composition with and without oil encapsulated in nanofibers, it was found that the 
CO – 50:50 sample with encapsulated oil showed higher digestibility than the other feed samples. For the other 
compositions, all feed samples without encapsulated oil exhibited higher digestibility compared to the formulations 
with encapsulated oil. When comparing the different feed compositions in intestinal fluid, the highest digestibility 
was observed for the CO – 100:0 sample, while the lowest digestibility was recorded for the CO – 50:50 sample. 
Among the feeds with encapsulated oil, the highest digestibility in intestinal fluid was also observed for the CO – 
50:50 sample. 

 



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Figure 4. Comparison between fish feed and feed containing nanofibers in vitro digestibility. The test was performed maintaining the 
samples in simulated gastric fluid for 1 hour (A) the test was performed maintaining the samples in simulated intestinal fluid for 5 hours (B). 
Note: Results followed by different capital letters differ significantly between the different treatment (fish feed a fndeed containing nanofibers) of the same 

compositions (P<0.05).  Results followed by different lowercase letters differ significantly between the different compositions of the same treatment 
(P<0.05).  Two-way ANOVA and Tukey’s test were used to determine statistical significance. 

 

4 Discussions 
4.1. Scanning Electron Microscopy (SEM) 

The study of nanofibers morphology can understand the encapsulation process. The addition of oil mixtures 
led to an increase in the average diameter of nanofibers (Table 2), which can be attributed to an increase in the 
polymeric solution viscosity. The relationship between viscosity and the average diameter of nanofibers was 
already noted in several papers [19, 21]. This behavior indicates that solutions containing different fractions of 
vegetable oils are more viscous, creating fibers with higher diameter values. 

 

4.2. Fourier-Transform Infrared Spectroscopy (FTIR) 
Infrared characterization enables the analysis of intermolecular interactions between the polymer and the 

active compounds through the identification of their chemical groups. The peaks observed for the polylactic acid 
nanofibers, corn oil, and gold linseed oil in the region from 3100 to 2800 cm-1 of the infrared spectrum agree with 
what is described in the literature [26, 33, 34]. The peaks show in the infrared spectrum region that confirm the 
encapsulation of vegetable oils in the polylactic acid matrices because it shows two signature peaks of corn and gold 
linseed oils (at 2925 cm-1, relative to CH3 symmetric stretch and at 2853 cm-1, relative to stretching vibration of 
bonds between carbon (C) and hydrogen (H) (CH stretch) in the nanofibers [33-36]. 

The FTIR results (Figure 2) suggest that weak interaction occurs between the gold linseed oil, corn oil, and 
the polylactic acid macromolecules occurs since there is no evidence of the formation of new primary bonds that 
can be associated with the shift or formation of new peaks in the infrared spectrum. Rahmani, et al. [33] observed 
alterations in the infrared spectrum of silicon nanowires (SiNWs) after the deposition of poly(3-hexylthiophene) on 
its surface. The formation of new peaks related to stretching vibration of bonds between carbon (C) and hydrogen 
(H) (C-H) and silicon (Si) and carbon (C) vibrations (Si-C) and the absence of the carbon (C) and carbon (C) (C=C) 
bond peak evidenced the poly(3-hexylthiophene) adsorption in the SiNWs surface through the formation of Si-C 
bonds after the rupture of the double bond between carbons. Another example is the paper of Yu, et al. [34], in 
which a shift of characteristic peaks of polyacrylonitrile nanofibers to a lower wavenumber after the encapsulation 
of different concentrations of acyclovir in the polymeric matrix, indicating the formation of hydrogen bonds 

between the carbon (C) and Nitrogen (N) (C≡N group) from polyacrylonitrile and the Oxygen (O) and hydrogen 
(H) (–OH group) from acyclovir. 

The presence of characteristic peaks of polylactic acid and vegetable oils in the nanofibers is an indication that 
the Solution Blow Spinning process was efficient to encapsulate these nutrients. 

 

4.3. Contact Angle 
It was expected that nanofibers would exhibit a hydrophobic behavior, with contact angle greater than 90° 

(Table 3), because both polylactic acid and vegetable oils are nonpolar molecules and, therefore, are incapable of 
forming intermolecular interactions with water. Moreover, the interaction between a polar liquid and a nonpolar 
surface is weak since the interactions inside the liquid are favored, resulting in a decrease of contact with the 
surface [36]. 

Paragkumar, et al. [37] showed that PLA films had a contact angle equal to 96°. This shows that the 
production of materials in nanoscale can improve properties such as hydrophobicity [38]. 

Therefore, the results align with following what is described in the literature and it can be affirmed that all 
nanofibers produced in this paper have great potential to retain the diffusion of the oil fraction in aquatic 
environments since there will be no significant interaction between the polymeric matrix and water.  

 
 
 



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4.4. Encapsulation Efficiency 
Knowing that the encapsulation process was successful, it is necessary to quantify the number of active 

compounds that were trapped in nanofibers. The CO: LO – 50:50 nanofiber had the one with the lowest 
encapsulation efficiency, which indicates that this sample will present an initial burst more evident than the other 
formulations, as shown by Wang, et al. [39]. In general, these results agree with what is reported in the literature 
about encapsulation in nanofibers. In the paper of Wang, et al. [39] the encapsulation efficiency of rosemary 
essential oil in polylactic acid nanofibers obtained by electrospinning was 90.18±1.77%, whereas the same 
encapsulation in pellets obtained by the extrusion technique led to an encapsulation efficiency of 68.77±1.41%. This 
behavior was also observed when comparing these two projects: Pérez-Masiá, et al. [40] obtained an encapsulation 
efficiency of 44.0±5.5% when encapsulating folic acid in starch microcapsules (electrospraying technique), and 
Fonseca, et al. [41] obtained an encapsulation efficiency of 95.6±0.0% when encapsulating folic acid in starch 
nanofibers (electrospinning technique). The higher values of encapsulation efficiency observed to nanofibers when 
compared with other polymeric matrices is due to properties like high surface-to-volume ratio, which reduce the 
number of interest compounds that can be present in the material surface. 

Through the comparison between the values demonstrated in Table 2 and Table 4, it can be assumed that the 
diameters of the nanofibers do not interfere directly in the encapsulation process, since large diameters are not 
necessarily associated with higher encapsulation efficiency values. 

 

4.5. Assessment of Nutrient Liberation in an Aqueous Medium (In Vitro) 
In fish farming, the feed does not stay in contact with water for more than 10 minutes [42] and through 

analysis of Figure 3, it is notable that in this period all nanofibers released between 14-21% of the nutrients 
encapsulated, while all feed samples released between 14-51% of nutrients during the same time. These results 
show that nanofibers are more effective in protecting nutrients against premature release.  

The nutrient release from nanofibers in the first 10 minutes of contact with an aqueous medium (Figure 3) has 
a direct connection with the high amount of vegetable oils encapsulated (50% w/w). This behavior was already 
explained by Karuppuswamy, et al. [43] who noticed that greater active agent concentrations led to faster burst 
release because there was a higher quantity of the compounds deposited in the polymeric matrix surface that could 
be easily dispersed in the medium [44]. The higher percentage of nutrients release observed for the feed can be 
explained by the fact that the feed commonly used in fish farming has approximately 40 essential nutrients that 
have to be present in specific quantities to ensure a balanced diet [45]. Since the formulation is extremely complex 
and composed of several hydrophilic and hygroscopic fractions, like soybean meal, cornflour, and dicalcium 
phosphate, a fast erosion of the feed granules occurs when in contact with water. This rapid erosion leads to an 
accelerated migration of the apolar fraction of nutrients, for example, vegetable oils. Therefore, they are dispersed 
in the aqueous medium before ingestion by fish. Notably, nanofibers were capable of retain the action compounds 
much more efficiency than the feed containing free oils. These results prove that the encapsulation of active agents 
was successful and confirms that the proposal to apply nanofibers in fish farming to feed fortification is promising 
since lower release in water is directly associated with greater availability of nutrients that can be absorbed in fish 
metabolism. 

 

4.6. Assessment of in Vitro Digestibility 
The fish feeding process starts when the feed comes into contact with water. At this moment, part of the 

nutrients added in the formulation is lost to the aquatic medium. Then, the pellets are consumed by the fish and the 
fraction of the active agents that were not released in water are digested in the stomach and, subsequently, in the 
intestine.  

The analysis of Figure 3 and 4 show the absence of a release standard between the feed samples, which 
indicates that the formulation presents certain heterogeneity and that the chemical composition of the oil fraction 
added in the feed has a direct impact on the apolar nutrients release since there was a big difference between the 
percentage of nutrients released in the simulated mediums for each feed sample. 

 Therefore, considering the heterogeneity of the formulation and the loss of feed nutrients that occurs in water, 
each pellet has different quantities of active compounds when they reach the gastrointestinal tract. This leads to 
consequences in the growth, development, and nutritional value of fishes since each one will consume different 
concentrations of nutrients. The final weight variation of farmed fishes that are fed with the conventional feed has 
been already observed by several authors, such as [46-48]. 

Nanofibers showed a more significant release in the simulated intestinal fluid than in simulated gastric fluid, 
which is a promising result since nutrients stay longer in the intestine, where most of its absorption occurs [16]. 
Furthermore, statistical analysis shows that even with different encapsulation efficiency and average diameter 
values, nanofibers can have similar effectiveness. 

The sample with the lowest encapsulation, efficiency (CO: LO – 50:50) was the one that released the greatest 
number of vegetable oils in water (Figure 3), which makes sense if we think that the lower the quantity of nutrients 
encapsulated the greater the fraction that is free in the polymeric matrix surface. The sample with the highest 
encapsulation efficiency (CO: LO – 100:0) was the one that released the greatest amount of vegetable oils in the 
intestinal fluid (Figure 4b), indicating that the polymeric matrix effectively protects the active agents, ensuring a 
pH-induced release.  Since the feed components are not wrapped by a protective layer, nutrients are free and 
released indiscriminately both in the aqueous medium and in the simulated gastrointestinal fluids. Thus, the feed 
contains a small number of nutrients when reaches the intestinal fluid, which affects the growth and development 
of fishe. Therefore, vegetable oil encapsulation in nanofibers can be extremely advantageous, not only 
environmentally but also economically and nutritionally. 

 

5. Conclusion 
The results showed that the encapsulation of vegetable oils in polylactic acid nanofibers obtained by the 

Solution Blow Spinning technique was efficient, and the product can be applied as a controlled release system to 



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improve fish nutrition and reduce the environmental impact associated with aquaculture. However, future 
experiments are needed to verify the efficiency of the feed containing encapsulated oil on the performance and 
physiology of the animals. 

 

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https://doi.org/10.1007/s11095-010-0239-y
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