Revista de Biología Tropical, ISSN: 2215-2075, Vol. 72(S1): e58995, marzo 2024 (Publicado Mar. 01, 2024)

Fatty acid composition and metabolic pathways in Arbacia dufresnii

(Arbaciidae: Arbacioida) gametes: implications of shrimp

byproducts in aquaculture feeds

Mercedes Vera Piombo1, 2; https://orcid.org/0000-0002-0361-7845

Marisa Avaro2; https://orcid.org/0000-0002-4923-8189

Agustín Gittardi2; https://orcid.org/0009-0004-3980-9031

Maximiliano Cledon3; https://orcid.org/0000-0002-1757-7274

Tamara Rubilar1, 2*; https://orcid.org/0000-0003-1728-3273

1. Laboratorio de Oceanografía Biológica (LOBio), CESIMAR- CCT CENPAT CONICET. Bv. Brown 2915, 9120U, Puerto

Madryn, Chubut, Argentina; mpiombo@cenpat-conicet.gob.ar, rubilar@cenpat-conicet.gob.ar (*Correspondence)

2. Laboratorio de Química de Organismos Marinos (LABQUIOM), Instituto Patagónico del Mar (IPAM), Facultad de

Ciencias Naturales y Ciencias de la Salud, Universidad Nacional de la Patagonia San Juan Bosco, Bv. Brown 3051,

9120U, Puerto Madryn. Chubut, Argentina; mavaro@unpata.edu.ar; aagittardi@gmail.com

3. Centro de Investigación aplicada y transferencia tecnológica en recursos Marinos “Almirante Storni”, Güemes 1030,

San Antonio Oeste, Argentina; mcledon@gmail.com

Received 28-VI-2023. Corrected 19-XII-2023. Accepted 08-I-2024.

ABSTRACT

Introduction: Care towards nutrition is essential for the quality of a sustainable aquaculture product. Since the

balance in food affects the growth and production of gametes. The circular economy is made possible through

the use of discarded materials.

Objective: The aim of this research was to study the fatty acid composition and metabolic pathways in the gam-

etes of Arbacia dufresnii, with a focus on the implications of incorporating shrimp byproducts into aquaculture

feeds.

Methods: Four different treatments were designed to maintain optimal nutritional quality, particularly in lipids

and proteins, based on previous studies. The fatty acid profiles of the feeds and gametes were analyzed by using

gas-chromatography, and statistical analyses were conducted to determine significant differences.

Results: Significant differences were observed in the abundance (%) of omega-3 (ω-3) and omega-6 (ω-6) fatty

acids. The (ω-3) metabolic pathway was more pronounced in the gametes of wild animals and those fed with the

experimental feeds. In contrast, the (ω-6) metabolic pathway was less relevant in these groups. The (ω-3) /(ω-6)

ratio was highest in the gametes of wild animals. Feeds enriched in fatty acids enhanced their bioaccumulation

in the gametes reaching higher concentrations than wild animals. The availability of fatty acids in foods allowed

their bioaccumulation in gametes, with concentrations equal to or higher than those observed in animals in their

natural environment for certain fatty acids.

Conclusions: Incorporating shrimp byproducts in aquaculture feeds demonstrated a promising strategy for

resource utilization and organic input generation. The fatty acid composition in the gametes of A. dufresnii was

influenced by the diet, highlighting the potential of balanced feeds to enhance the bioaccumulation of essential

fatty acids. These findings provide valuable insights for the development of sustainable aquaculture practices and

the production of nutritionally enriched seafood products.

Key words: sea urchin; reproductive cells; lipid profile; biochemical pathways; marine byproducts; sustainable

feeds.

https://doi.org/10.15517/rev.biol.trop..v72iS1.58995

SUPPLEMENT

2Revista de Biología Tropical, ISSN: 2215-2075 Vol. 72(S1): e58995, marzo 2024 (Publicado Mar. 01, 2024)

INTRODUCTION

Sea urchin aquaculture has shown great

potential for growth in recent years (Rubilar

& Cardozo, 2021) with research focused on

the development of feed formulations that can

optimize growth and gonad development in

commercially available species (Lawrence et al.,

2013). Formulated dry feeds have become an

essential component of aquaculture practices

for marine species, offering several advantages

over traditional feeds such as consistency in

quality and composition, water stability, and

ease of use. The use of formulated feeds in sea

urchin aquaculture is crucial for the establish-

ment of large-scale production facilities and the

commercial viability of the industry. However,

the nutritional content and ingredient quality

of the feeds play a significant role in the success

of sea urchin aquaculture, with research indi-

cating that specific feed formulations can have

a significant impact on gonad development

(Martínez-Pita et al., 2010; Raposo et al., 2019).

In addition to the importance of feed qual-

ity, feed costs can represent a significant por-

tion of the total production cost in aquaculture.

To address this issue, feeds formulated with

viable nutritional levels of by-products have

been developed, reducing production costs and

contributing to environmental sustainability

(Vizzini et al., 2019; Ciriminna et al., 2021). By-

product feeds, such as shrimp and squid meals,

have been successfully used as ingredients for

sea urchin feed formulations, improving feed

efficiency and reducing organic waste gener-

ated in the fish and seafood processing industry

(Gunathilaka et al., 2021). These developments

have contributed to the circular economy, pro-

moting the sustainable use of resources and

reducing waste.

Shrimp is an important economic

resource in the Argentinean Patagonian region

RESUMEN

Composición de ácidos grasos y vías metabólicas en los gametos de Arbacia dufresnii

(Arbaciidae: Arbacioida) implicaciones de los subproductos de langostinos en los alimentos de acuicultura

Introducción: El cuidado hacia la nutrición es fundamental para la calidad de un producto acuícola sostenible. Ya

que el balance en los alimentos afecta el crecimiento y producción de los gametos. A partir del aprovechamiento

de materias de descarte se posibilita la economía circular.

Objetivo: El objetivo de este estudio fue investigar la composición de ácidos grasos y las vías metabólicas en los

gametos de Arbacia dufresnii, centrándose en las implicaciones de la incorporación de subproductos de camaro-

nes en los alimentos de acuicultura.

Métodos: Se diseñaron cuatro tratamientos diferentes para mantener una calidad nutricional óptima, especial-

mente en lípidos y proteínas, basándose en estudios previos. Se analizaron los perfiles de ácidos grasos de los

alimentos y los gametos mediante cromatografía de gases, y se realizaron análisis estadísticos para determinar

diferencias significativas.

Resultados: Se observaron diferencias significativas en la abundancia (%) de ácidos grasos omega-3 (ω-3) y

omega-6 (ω-6). La vía metabólica de (ω-3) fue más pronunciada en los gametos de los animales en su entorno

natural y aquellos alimentados con los piensos experimentales. Por el contrario, la vía metabólica de (ω-6) tuvo

menos relevancia en estos grupos. La relación (ω-3) /(ω-6) fue más alta en los gametos de los animales en su

entorno natural. La disponibilidad de ácidos grasos en los alimentos permitió su bioacumulación en los gametos,

con concentraciones iguales o superiores a las observadas en los animales en su entorno natural para ciertos

ácidos grasos.

Conclusiones: La incorporación de subproductos de camarones en los alimentos de acuicultura demostró ser una

estrategia prometedora para la utilización de recursos y la generación de insumos orgánicos. La composición de

ácidos grasos en los gametos de A. dufresnii fue influenciada por la dieta, destacando el potencial de los alimentos

balanceados para mejorar la bioacumulación de ácidos grasos esenciales. Estos hallazgos brindan información

valiosa para el desarrollo de prácticas sostenibles en acuicultura y la producción de productos marinos enrique-

cidos nutricionalmente.

Palabras clave: erizo de mar; células germinales; perfil lipídico; vías metabólicas; subproductos marinos; alimen-

tos sostenibles.

Revista de Biología Tropical, ISSN: 2215-2075, Vol. 72(S1): e58995, marzo 2024 (Publicado Mar. 01, 2024)

(Bondad-Reantaso et al., 2012). Shrimp pro-

cessing generates large amounts of waste,

including heads, shells, and other by-products,

which can be a source of environmental pollu-

tion if not properly managed. In Patagonia, for

instance, the shrimp industry generates an esti-

mated 60 000 tons of waste per year (González-

Zevallos et al., 2020; Moriondo & de la Garza,

2020). In this context, the development of

shrimp-based feed formulations has the poten-

tial to contribute to both the economic and

environmental sustainability of the region.

Several studies have investigated the use of

shrimp waste as a source of protein and other

nutrients for the production of aquaculture

feeds (Abuzar et al., 2023; Canseco et al., 2015;

Espinosa-Chaurand et al., 2015). For example,

Espinosa-Chaurand et al. (2015), formulated a

shrimp-based feed that achieved similar growth

rates and feed conversion ratios to those of

a commercial feed. Similarly, Canseco et al.

(2015) found that shrimp waste could replace

up to 50 % of fishmeal in the diet of juve-

nile white shrimp. Shrimp meal has also been

used as a protein source in sea urchin feed,

with studies showing that it can improve feed

efficiency and reduce the cost of feed. These

findings suggest that the use of shrimp waste in

feed formulations may reduce the cost of feed

production while contributing to the circular

economy by reducing waste.

Although much research has been con-

ducted on the effect of feed on gonads in com-

mercially available sea urchin species, little

is known about the effect of feed on Arbacia

dufresnii (Blainville, 1825) a novel commercial

species (Lawrence et al., 2013). This species is

attractive to consumers due to the production

of nutraceutical products manufactured by a

Start Up EriSea S.A. One of the products has

high levels of polyunsaturated fatty acids and

high polyunsaturated fatty acids (PUFAs and

HUFAs) from their gonads (Díaz de Vivar et al.,

2019). Since PUFAs and HUFAs are important

for human health and have been shown to have

anti-inflammatory, anti-cancer, and cardio-

vascular benefits (Jobling, 2012; Simopoulos,

2010; Trenzado et al., 2012) and the fatty acid

composition of sea urchin gonads and eggs can

be influenced by feed quality and composition

(David et al., 2020; Kozhina et al., 1978); it is

important to investigate the effect of feed on the

fatty acid composition of A. dufresnii gonads.

The most important HUFA are docosa-

hexaenoic acid (DHA) and Arachidonic acid

(ARA), since they are components of cell mem-

brane phospholipids and play important roles

in cell division, differentiation, and signaling

(National Research Council, 2011). Besides

eicosapentaenoic acid (EPA), which is also an

essential HUFA, is present in high propor-

tions in marine organisms; (Bell et al., 2003).

Therefore, the incorporation of HUFAs in the

diet of A. dufresnii may have a positive impact

on gonad development and fatty acid composi-

tion. Therefore, this study aims to evaluate the

effect of feed on the fatty acid composition of A.

dufresnii eggs. Understanding how feed affects

the fatty acid composition of this novel com-

mercial species can provide valuable informa-

tion for the development of feed formulations

that optimize growth and gonad development,

and increase the commercial viability of sea

urchin aquaculture.

MATERIALS AND METHODS

Specimens’ collection: Mature individu-

als of the species A. dufresnii were collected by

scuba diving during the winter and spring of

2021, as well as at the end of summer in 2022.

The collection took place in Puerto Madryn

(42º 43.6” S & 65º 1.2” W), Gulf Nuevo Bay,

Northern Patagonia, Chubut, Argentina. Sub-

sequently, the individuals were transported in

seawater to the Erisea S.A. pilot aquaculture

facility. Once there, the individuals were sorted

by sex through the induction of spawning using

an injection of KCl [0.5 M]. The individuals

were then kept in a fasting state for a minimum

period of five days after spawning.

Experimental design: Four different fatty

acids concentrations in feed were tested. Their

concentrations in gametes were analyzed before

and after 60 days. A total of 2 240 females

4Revista de Biología Tropical, ISSN: 2215-2075 Vol. 72(S1): e58995, marzo 2024 (Publicado Mar. 01, 2024)

were separated into the four treatments, each

treatment with four replicates (140 individuals

per replicate).

Individuals of A. dufresnii were randomly

distributed into baskets within a production

tank. Throughout the 60-day duration of the

experiment, environmental conditions were

kept constant, including temperature (14 ± 2

ºC) and photoperiod (12:12 h) (Sepúlveda et al.,

2021), and periodic monitoring of ammonia,

nitrite, nitrate, dissolved oxygen, and salinity

values (35 ppm) was conducted to maintain

them within recommended ranges (Rubilar &

Crespi-Abril, 2017). The animals were fed ad

libitum with 400 mg feed per individual every

three days, following the protocol described by

(Rubilar et al., 2016).

Feed: Four balanced feeds were developed

with different concentrations of fatty acids for

aquaculture. Each diet shared 76 % of its basic

formulation, while the remaining 24 % was

differentiated by the addition of flours from

different parts of Pleoticus muelleri (Spence

Bate, 1888) shrimp: pulp (TA), cephalothorax

(TD), or a combination of both with exoskel-

eton (TB or TC; 12 %: 12 % respectively). The

formulation of each diet was based on theo-

retical nutritional information of the ingredi-

ents obtained from an aquaculture formulators’

database (Table 1) and the concentration of

essential fatty acids (Table 2). Furthermore,

analyses were conducted on the feeds to verify

the actual FA concentration in each ingredient.

Gametes collection: The oocytes of A.

dufresnii females were collected before the start of

the experiment and at its conclusion, after being

fed with the treatments. They were induced to

spawn by injecting 0.3 mL of KCl [0.5 M] into

the peristomal membrane (Sun & Chiang, 2015).

Once injected, the females were placed in direct

contact with previously filtered and UV-treated

seawater, and a pool of oocytes was collected

from each experimental replicate. Subsequently,

the oocyte sample was allowed to settle in a

Table 1

Proximal composition (%) for each feed.

Nutrient (%) TA TB TC TD

Lipids 4.17 4.83 5.47 5.49

Soluble protein 12.18 15.86 17.67 19.55

Insoluble protein 16.48 21.10 23.04 25.73

Fiber 7.05 8.56 10.80 10.07

Ashes 15.96 18.94 22.46 21.91

Abbreviations from balanced feeds treatments’ labeled: TA, 24 % of P. muelleri pulp; TB, combination pulp and exoskeleton

of P. muelleri (12 %: 12 %); TC, combination cephalothorax and exoskeleton of P. muelleri (12 %: 12 %); TD, cephalothorax

of P. muelleri (24 %).

Table 2

Specific differentiation in the composition of fatty acids in feed.

Fatty acids (%) in feed TA TB TC TD

LA 1.66 1.58 1.51 1.51

ALA 0.62 0.52 0.44 0.41

ARA 0.30 0.31 0.30 0.32

EPA 0.36 0.99 1.90 1.62

DHA 2.56 2.14 1.96 1.72

Abbreviations from balanced feeds treatments’ labeled: TA, 24 % of P. muelleri pulp; TB, combination pulp and exoskeleton

of P. muelleri (12 %: 12 %); TC, combination cephalothorax and exoskeleton of P. muelleri (12 %: 12 %); TD, cephalothorax

of P. muelleri (24 %). Fatty acids abbreviated names: LA, Linoleic acid; ALA, α-Linolenic acid; ARA, Arachidonic acid; EPA,

Eicosapentaenoic; DHA, Docosahexaenoic acid.

Revista de Biología Tropical, ISSN: 2215-2075, Vol. 72(S1): e58995, marzo 2024 (Publicado Mar. 01, 2024)

settling funnel for a maximum of 12 h. at 4 ºC.

Ten milliliters of oocytes from each treatment

were stored at -20 ºC under the N2 atmosphere

until further processing, while maintaining light

protection throughout the process.

Fatty acid (FA) determination: Gametes

from A. dufresnii females, as well as meal

from P. muelleri and the four feeds used in

the experiment, were employed. To obtain the

FAME (fatty acid methyl ester), a 100 mg dry

sample (dried at 40 ºC) was taken and sub-

jected to transmethylation using the Lepage

& Roy (1986) method, with tricosanoic acid

as the standard (51.04 μg). The separation

and quantification of FAME were performed

using gas chromatography with mass spectrom-

etry detection (GC-MS) on a Thermo Focus

ISQ instrument. Peaks corresponding to each

detected FAME molecule were identified by

comparing their relative retention times with

authentic standards (a mixture of 37 FAME

species from Supelco 47885-U). Additionally,

mass spectra were analyzed and compared to

the NIST library (National Institute of Stan-

dards and Technology, USA) and The Lipid

Web database for confirmation (Christie, 1998;

Fahy et al., 2007).

The concentration of fatty acids (FA) was

calculated in the four formulated feeds and

A. dufresnii gametes. The values are present-

ed as mean (± SD) in μg·g-1 of dry sam-

ple. FA were expressed using their I.U.P.A.C

nomenclature and corresponding abbreviated

names, which include: Linoleic acid (LA, C18:2

(n-6)), α-Linolenic acid (ALA, C18:3 (n-3)),

Arachidonic acid (ARA, C20:4 (n-6)), Eicosa-

pentaenoic acid (EPA, C20:5 (n-3)), and Doco-

sahexaenoic acid (DHA, C22:6 (n-3)). The FA

were grouped according to the abundance (%)

of the nutritional metabolic pathway in (ω-3)

(ALA→EPA→DHA) and (ω-6) (LA→ARA). The

(ω-3) /(ω-6) ratio was calculated.

Statistical analysis: Analysis of variance

(ANOVA) was used to test the significance of

the concentration of each FA and the abun-

dance (%) grouped according to the metabolic

pathways, (ω-3) and (ω-6), in the gametes of A.

dufresnii females from the treatments, includ-

ing the wild animals. The assumptions of nor-

mality and homogeneity of variances were

verified using the Welch method, α = 0.05 (©

Minitab, 2017), and were checked before all

ANOVA analyses.

RESULTS

Fatty acids concentration: Based on the

data obtained from GC-MS chromatography,

the concentration was calculated for the FA

in the four food types (TA, TB, TC, TD). TA

showed the highest concentration of the stud-

ied FAs, being 50 times richer in LA, 60 times

richer in ALA, EPA, and DHA, and 11 times

richer in ARA. Although treatments TB, TC,

and TD have similar concentrations among

themselves, there is a decreasing trend in the

concentration of FAs, especially in LA. The

concentration of FAs, expressed as (μg·g-1), in

the four foods (TA, TB, TC, TD) is detailed in

Table 3.

Table 3

Fatty acids concentration (μg·g-1) in the four foods.

Feed LA ALA ARA EPA DHA

TA 298 865.46 92 441.77 9 131.53 78 770.08 88 959.84

TB 5 118.59 1 642.88 837.83 1 331.61 1 518.34

TC 5 014.57 1 620.38 48.01 1 358.97 1 591.51

TD 4 707.16 1 476.31 169.66 1 251.24 1 455.72

Abbreviations from balanced feeds treatments’ labeled: TA, 24 % of P. muelleri pulp; TB, combination pulp and exoskeleton

of P. muelleri (12 %: 12 %); TC, combination cephalothorax and exoskeleton of P. muelleri (12 %: 12 %); TD, cephalothorax

of P. muelleri (24 %). Fatty acids abbreviated names: LA, Linoleic acid; ALA, α-Linolenic acid; ARA, Arachidonic acid; EPA,

Eicosapentaenoic; DHA, Docosahexaenoic acid.

6Revista de Biología Tropical, ISSN: 2215-2075 Vol. 72(S1): e58995, marzo 2024 (Publicado Mar. 01, 2024)

The fatty acids composition: FA composi-

tion in the female gametes presented significant

differences among treatments. They were also

different from the wild individuals analyzed in

this study (Fig. 1). TA and TB exhibited higher

values of LA (5 026 ± 435 μg·g-1; 4 903 ± 1 199

μg·g-1), respectively, in the diets. On the other

hand, from wild animals’ values showed the

lowest FA concentration (1 346 ± 468 μg·g-1) (F

(4, 21; 0.05) = 19.3, P = 0.000). TC had the lowest

values of ALA (1 142 ± 301 μg·g-1), similar to

the values found in wild gametes (1 192 ± 374

μg·g-1) (F (4, 21; 0.05) = 5.3; P = 0.005). Although

the ARA values in TC seem to be the highest

(2 532 ± 1 764 μg·g-1), there were no significant

differences (1 496 ± 471 μg·g-1) (F (4, 21; 0.05) =

2.57; P = 0.073). TD had the highest values of

EPA (4 909 ± 816 μg·g-1) among the treatments,

which were very similar to the wild gametes

(5 256 ± 729 μg·g-1), however, no significant

differences were observed (4 541 ± 946 μg·g-

1) (F (4, 21; 0.05) = 1.66; P = 0.203). TA and TB

exhibited the highest values of DHA (2 643 ±

261 μg·g-1; 2 479 ± 39 μg·g-1), respectively (F (4,

21; 0.05) = 14.64; P = 0.000).

Fatty acids abundance: There were signifi-

cant differences in the abundance (%) of both

(ω-3) and (ω-6). The (ω-3) metabolic pathway

was higher in the gametes of wild animals

and those fed with TD (F (4, 21; 0.05) = 8.56; P =

0.000). On the other hand, the (ω-6) metabolic

pathway was less relevant in this group (F (4, 21;

0.05) = 8.56; P = 0.000) (Fig. 2). The (ω-3) /(ω-6)

ratio showed the highest values in the gametes

from wild animals (F (4, 21; 0.05) = 12.71; P =

0.000), with increasing values observed from

TA to TD treatments (Table 4), and the wild

had the highest ratio.

Fig. 1. Concentration of fatty acids (μg·g-1) in female gametes of A. dufresnii. A. gametes from treatment A; B. gametes from

treatment B; C. gametes from treatment C; D. gametes from treatment D; and gametes from Wild animals spawning before

treatments.

Table 4

The (ω-3) /(ω-6) ratio of the abundance (%) in gametes of

A. dufresnii.

Feed TA TB TC TD Wild

Ratio 1.43 1.44 1.52 2.10 3.37

Abbreviations from balanced feeds treatments’ labeled:

TA, 24 % of P. muelleri pulp; TB, combination pulp and

exoskeleton of P. muelleri (12 %: 12 %); TC, combination

cephalothorax and exoskeleton of P. muelleri (12 %: 12 %);

TD, cephalothorax of P. muelleri (24 %).

Revista de Biología Tropical, ISSN: 2215-2075, Vol. 72(S1): e58995, marzo 2024 (Publicado Mar. 01, 2024)

DISCUSSION

In this study, the female gamete fatty acid

composition was studied using different con-

centrations of fatty acids in the feed. The

origin of the fatty acid were different parts of

the shrimp P. muelleri. The cephalothorax was

identified as the most abundant part in (ω-3)

fatty acids, particularly EPA and DHA (Cretton

et al., 2020; Hop et al., 1990). Conversely, the

exoskeleton is considered a by-product of the

shrimp industry, resulting from the processing

for human consumption (De Carli et al., 2012).

The utilization of this shrimp waste in the feed

provided to A. dufresnii demonstrates an inter-

esting strategy for resource utilization and the

generation of organic inputs in aquaculture.

The feed composition used in this study is

highly complex, incorporating ingredients from

diverse sources such as terrestrial vegetable

meals, fish oil, and algae meal. This complexity

may impact the bioavailability and potential

oxidation of fatty acids during the pelletization

process of the feeds. However, it is noteworthy

that LA and ALA are essential fatty acids pri-

marily originating from terrestrial plant sources

(Prato et al., 2018), while ARA, EPA, and DHA

are predominantly derived from marine sourc-

es. These findings align with previous research

analyzing the fatty acid composition in differ-

ent feeds with varied nutrients that are better

assimilated (Espinoza et al., 2022; Fernandez

& Boudouresque, 1998; Lawrence et al., 2013;

Spirlet et al., 2001).

The different fatty acid composition in

the feed fed to A. dufresnii revealed that the

composition of the feeds used in the treatments

maintained is in an appropriate range of pro-

teins; considered of high nutritional quality for

echinoderms (Wilson, 2003). These findings

are consistent with previous research highlight-

ing the importance of maintaining adequate

nutritional quality in the diet of sea urchins

(Lawrence et al., 2013).

The availability of fatty acids in the feeds

allowed for their bioaccumulation in the gam-

etes, reaching concentrations equal to or higher

than those observed in wild animals for LA,

ALA, and DHA. ARA and EPA concentrations

remained stable. Our results indicate that the

duration of the 8-week experiment and the

selection of feeds were appropriate for obtain-

ing values equivalent to those observed in ani-

mals in their natural environment.

Fig. 2. Abundance (%) of FA according to their metabolic pathway in A. dufresnii gametes. A. gametes from treatment A;

B. gametes from treatment B; C. gametes from treatment C; D. gametes from treatment D; and gametes from Wild animals

spawning before treatments.

8Revista de Biología Tropical, ISSN: 2215-2075 Vol. 72(S1): e58995, marzo 2024 (Publicado Mar. 01, 2024)

During the gametogenesis stage, there is

an increase in the accumulation of (ω-3) fatty

acids, particularly EPA and ARA, in the gametes

of wild animals (Rahman et al., 2014; Rocha et

al., 2019; Sanna et al., 2017; Wang et al., 2020;

Zárate et al., 2016). Diet plays a significant role

in the bioavailability of these fatty acids, and it

has been demonstrated that balanced feeds can

enhance the bioaccumulation of EPA and DHA

in gametes studies investigating fatty acid con-

centrations in gametes have reported elevated

levels of EPA and ARA in wild animals (Díaz

de Vivar et al., 2019; García & Pita, 2010; Qi et

al., 2018); and studies of modulation in the FA

accumulated in the eggs (Carboni et al., 2013;

Zhou et al., 2011). The availability of fatty acids

in the feeds allowed bioaccumulation in the

gametes, reaching concentrations equal to or

higher than those observed in wild animals for

LA, ALA, and DHA. ARA and EPA concentra-

tions remained stable in A. dufresnii. Therefore,

this study demonstrated that diet modulated

the fatty acid composition in female A. dufresnii

gametes.

The effect of overfeeding in the aquarium,

as mentioned by Guillou et al. (2000), is a

phenomenon that occurs when animals have

low energy expenditure and accumulate excess

energy in their tissues. The excessive contribu-

tion of fatty acids from the TA diet, compared

to LA and ALA, may be related to this over-

feeding effect, which was reflected in a dis-

proportionate accumulation of EPA and DHA.

Therefore, the formulation of balanced diets

and regulation of dosage to meet the nutritional

needs of A. dufresnii is crucial to avoid imbal-

ances in fatty acid metabolism.

In conclusion, this study demonstrated

significant differences in fatty acid composi-

tion and metabolic pathways in A. dufresnii

gametes under different feeding treatments.

Diets rich in (ω-3) fatty acids had a notable

effect on the (ω-3) metabolic pathway, while

diets rich in (ω-6) fatty acids deviated sig-

nificantly from the natural metabolic pathways

of the organisms. The TD diet, formulated

with shrimp waste, resulted in gametes with

fatty acid values similar to those found under

natural feeding conditions, making it a suitable

option for maintenance diet in aquaculture.

The study also highlighted the potential of uti-

lizing shrimp waste as a resource in aquaculture

and promoting the development of a sustain-

able blue economy.

Ethical statement: the authors declare that

they all agree with this publication and made

significant contributions; that there is no con-

flict of interest of any kind; and that we fol-

lowed all pertinent ethical and legal procedures

and requirements. All financial sources are fully

and clearly stated in the acknowledgments sec-

tion. A signed document has been filed in the

journal archives.

ACKNOWLEDGMENTS

We want to thank the technology-based

company Erisea S. A. for allowing the space and

equipment to carry out the experiment, as well

as the participation of the technical staff.

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