1.
INTRODUCTION:
Aquaculture is one of the fastest-growing sectors in
global animal protein production (Hoseini and Al Sulivany,
2024). In the aquaculture sector, silicon nanoparticles (Si-NPs) have gained
considerable interest due to their potential to boost disease resistance,
improve growth rates, and alleviate environmental challenges, including the
effects of heavy metal toxicity (Khalefa et al., 2024). Si-NPs play a potential
role in regulating immune functions by boosting the efficiency of antioxidant
enzymes, lowering oxidative stress, and enhancing general fish health (Mahboub et
al., 2024; Rahman et al., 2025). Additionally, these nanoparticles
have been shown to reduce the harmful effects of heavy metals and combat
pathogenic diseases, thereby supporting the sustainability and resilience of
aquaculture systems (Khalefa et al., 2024; Ali et al., 2024). Their
potential to enhance nutrient absorption and promote beneficial gut microbiota
further solidifies their role as an effective and promising feed supplement in
fish nutrition (Hussain et al., 2024; Ghafarifarsani et al.,
2024; Hamed & Badran, 2024; El-Naby et al., 2025).
In developing countries across Asia and Africa, the expansion of the
aquaculture sector plays a vital role in ensuring economic stability and
strengthening food security (Chan et al.,
2019). The effectiveness of aquaculture practices largely depends on
the quality of feed, which directly impacts production efficiency and economic
sustainability (Singha et al., 2021). The use of Si-NPs in aquafeeds has been linked to enhanced feed
efficiency and better growth outcomes in multiple fish species.
Heavy metal pollution in aquatic environments poses a serious risk
to fish health, leading to oxidative stress, weakened immune systems, and
impaired organ function (Banday et al., 2019).
Harmful metals like lead, cadmium, and mercury accumulate in fish
tissues, disrupting cellular balance and metabolic processes, which ultimately
hampers growth and weakens immune function (Balali-Mood et
al., 2021; Reda et al., 2025). Moreover,
incorporating Si-NPs into fish diets has been shown to revive antioxidant
defense systems by lowering oxidative stress markers and boosting the activity
of antioxidant enzymes in fish affected by trace element toxicity (Ro et al., 2025).
Furthermore, the immunomodulatory properties of Si-NPs have
garnered significant attention in aquaculture. Research suggests that including
Si-NPs in fish diets notably strengthen immune responses by increasing
immunoglobulin levels, boosting cytokine production, and upregulating
immune-related gene expression, ultimately enhancing disease resistance in fish (Ghafarifarsani et al., 2024; Hussain et al.,
2024). In common carp (C. carpio), the addition of Si-NPs has
also demonstrated beneficial impacts on gut health, nutrient absorption, and
overall growth performance, including improvements in body composition (Hamed and Badran, 2024). The
incorporation of Si-NPs into aquafeeds has proven effective in promoting fish
growth, optimizing enzyme activity, boosting immune system performance, and
counteracting the harmful effects of heavy metal toxicity. However, these promising results, further
research is necessary to determine the ideal dosage, assess long-term
physiological impacts, and explore potential interactions with other functional
feed additives. This study
aims to investigate the effects of dietary Si-NPs on growth performance, enzyme
activity, and metal accumulation in C. carpio (common carp),
providing valuable insights into the application of nanotechnology for
sustainable aquaculture practices.
2.
MATERIALS AND METHODS:
Fish and Rearing Condition
One hundred twenty healthy C. carpio juveniles of the same
age group, weighing 12.00±1.50 g and a total length of 8.50±1.20 cm, were
obtained from Tawakal Fish Hatchery, Punjab, Pakistan. The fish was transported
to the aquaculture laboratory of the Saline Water Aquaculture Research Center,
Fisheries Department. Before the experiment began, the fish was acclimated to
laboratory conditions for 15 days to allow physiological adaptation. To
eliminate potential external pathogens, the fish were disinfected with sodium
chloride (NaCl) solution (Sigma-Aldrich) for 1–2 minutes, as per the
disinfection protocol provided by Zhang et al. (2024). During the
acclimatization period, fish were given a standardized basal diet twice daily
at fixed intervals to provide the best nutrition and health conditions
(Laz-Figueroa et al., 2024). After acclimatization, fish were allocated
randomly into twelve glass aquaria, each measuring 120×40×50 cm³ with a
capacity of 100 liters.
Preparation
of Experimental Diets:
Purely activated, 100% natural silica nanoparticles (SiO2; Si-NPs)
of case number item 7631-86-9, which were purchased from Sigma Aldrich,
Pakistan. The Si-NPs were recovered by the use of ethanol and thus gave a size
distribution between 300 nm to 400 nm with an average active density of
2.08±0.21 g/cm³ is equivalent to approximately 3×10¹⁶ nanoparticles per
gram. Post-purchasing, the Si-NPs were prepared at the Fish Aquaculture
Laboratory, Saline Water Aquaculture Research Center, Fisheries Department, to
enhance their bioavailability and functional characteristics so that they could
be applied for fish nutrition study.
In Table 1, all the ingredients were ground into powders using a blender. Then,
the required quantities of fish meal corn, soybean barely, ascorbic acid,
premix, and Si-NPs were weighed with an electronic
digital weighing balance. The ingredients were then blended in a feed mixer for
10 min. After that, slowly, 15ml of distilled water was poured and mixed to
create a dough. This dough was then passed through a 2.0 mm diameter
hand-driven pelleting machine. Extrusion-produced pellets were overnight stored
at 45oC and then preserved at -18oC to be used later
(Ceccotti et al., 2019).
Experimental design:
The design of the experiment included four
dietary treatments, each with three replicates with 10 fish stockings per tank,
to minimize stress levels. The experimental diets were categorized and labelled
as GRP-1; 0 mg/kg (control) Si-NP, GRP-2; 1 mg/kg Si-NP, GRP-3; 2 mg/kg Si-NP,
and GRP-4; 3 mg/kg Si-NP inclusion levels respectively. The trial was fed for
90 days. For aeration to provide water quality, high-efficiency aquarium air
pumps (SunSun CT-402, power: 6 W, airflow: 4.0 L/min) and industrial-grade air
compressors (Hailea V-20, power: 50 W, airflow: 75 L/min; Hailea V-30, power:
60 W, airflow: 85 L/min; Resun LP-100, power: 180 W, airflow: 0.140 m³/min) was
utilized. The water quality parameters were traced daily in detail and adjusted
as and when required to maintain proper conditions for fish growth and
development. The pH, dissolved oxygen, and temperature values measured were 7.2
± 0.20, 6.8 ± 0.5 mg/L, and 18 ± 0.4°C, respectively, to create a stable and
controlled aquatic ecosystem for the experiment.
Growth Measurement
After rearing was completed, fish were fasted for 24 hours to
allow complete gut clearance. Then, the individual fish were weighed on a
high-accuracy digital balance (A&D HT-120) to record their final body
weight. Growth performance parameters like initial weight (IW), final weight
(FW), Weight gain (WG), feed intake (FI), and feed
conversion ratio (FCR) were determined as below (Al Sulivany et al.,
2024; Owais et al., 2024a. b; Abdulrahman and Al Sulivany, 2025).
WG (g) =FW -IW.
FI (g) = Total feed given (g)/ Number of
fish - duration of feeding.
FCR = Feed intake (g) / WG (g).
Determination of Enzyme
Activity Parameters
After weighing, the fish were anesthetized
using an MS-222 (tricaine methane sulfonate, 0.1 g. l-1,
Sigma-Aldrich, USA) to minimize stress during the procedure. Blood samples were
collected from the caudal vein using 5 ml syringes. The blood was then inserted
into the gel tube without anticoagulants and centrifuged at 1500 rpm for 5
minutes to separate the serum for enzymatic activity parameters.
Preparation of Muscle
and Heavy Metal Determination.
After blood sampling, five fish from each
pond were euthanatized with a quick cranial percussion to avoid causing any
undue stress. Under sterile conditions using sterile surgical scalpels, a
muscle tissue piece of about 2g (1×1 cm) was removed from the mid-dorsal
position of each fish. The tissue samples so collected were oven-dried at 80–85
°C for 24 hours to remove the moisture content. Following thorough drying, the
samples were cooled to room temperature and finely powdered using a clean
ceramic mortar and pestle to render them uniform (Ahmed & Hasan, 2019).
For heavy metal analysis, 0.5 g of the homogenized muscle sample
was precisely weighed and transferred into Teflon digestion vessels, followed
by the addition of 10 mL of concentrated nitric acid (HNO3, 69%).
The digestion process was performed in a Milestone ETHOS UP microwave digestion
system under controlled conditions: ramping to 180 °C for 5 minutes,
maintaining digestion at 180 °C for 10 minutes, and allowing a cooling phase of
6 minutes. The completely digested samples were then transferred to
acid-cleaned volumetric flasks and diluted to a final volume of 10 mL using
ultrapure deionized water. The prepared solutions were stored at 4 °C until
further analysis (Al-Imarah et al., 2024). To quantify heavy metals,
including Pb and Cd, Inductively Coupled Plasma-Optical Emission Spectroscopy
(ICP-OES) (PerkinElmer Avio 500) was utilized. The system was equipped with a
radial torch and an argon saturation assembly to enhance signal stability.
High-purity (99.999%) argon gas was used for plasma, auxiliary, and nebulizer
functions, with optimized gas flow rates of 15.0 L/min for plasma, 1.3 L/min for
auxiliary, and 0.58 L/min for nebulizer. The plasma generator's radio frequency
(RF) power was set at 1.45 kW, and the plasma vertical height was adjusted to 8
mm for optimal detection sensitivity. The analytical parameters included a
sample uptake time of 20 sec, a delay time of 5 sec, a rinse time of 10 sec, an
initial stabilization time of 12 sec, and 5 sec between replicate analyses. The
emission wavelengths were selected for their high sensitivity in detecting
trace metals: Na⁺ (589.592 nm), Co²⁺ (238.892 nm), Ni²⁺
(213.857 nm), Fe²⁺ (267.716 nm), Mg²⁺ (259.940 nm), Al³⁺
(285.213 nm), Zn²⁺ (220.353 nm), Pb²⁺ (327.395 nm), Cu⁺
(231.604 nm), Cr³⁺ (396.152 nm), and Cd²⁺ (260.568 nm). Before
sample analysis, the instrument was calibrated using certified multi-element
standards to ensure precision and accuracy.
Ethical
approval and consent:
All authors gave verbal informed consent for their participation.
The study's design and procedures were reviewed and approved by the Animal
Ethics Committee of the Institutional Research Committee under the Government
Department of Fisheries, Punjab, Pakistan, in compliance with ethical standards
(Code APF-129; 2023).
Statistical
Analysis
Statistical analysis was performed using GraphPad Prism 9. A
one-way analysis of variance (ANOVA) was conducted, followed by Dunnett’s post
hoc test to compare each experimental group with the control. Results are
presented as means ± standard error (SE).
3.
RESULTS:
The result of the study evaluated the growth performance
of common carp feds diets supplemented with varying levels of Si-NPs,
indicating clear differences among the groups (Table 2 and Figure 1; A, B, C,
D, and E). The FW was highest in the GRP3 at 35±0.44 g, followed by GRP2 at 30.2±0.37 g, the GRP1 group at 25.8±0.37 g, and the GRP4 group at
22.8±0.33 g. Similarly, WG was
greatest in GRP3 (22.4 g), significantly higher than in GRP2, GRP1, and GRP4.
Feed intake (FI) was also highest in GRP3, while GRP4 had the lowest intake
(18.34 g). The FCR was most reduced in GRP3 (1.269), whereas GRP4 showed the
least efficiency (1.646). Statistical analysis confirmed significant
differences (P < 0.05) in FW, WG, FI, and FCR among the groups.
4.
DISCUSSION:
The experiment was conducted to evaluate
the effects of Si-NPs on growth, heavy metals, and serum enzyme activity of C.
carpio throughout the 90-day feeding trial. The current study confirmed
that Si-NPs supplementation at 1 mg/kg (GRP2) and 2 mg/kg (GRP3) enhanced the
growth performance of C. carpio significantly compared to the control
group (GRP1). The growth parameters exhibited significant differences among the
different treatment groups. Gao and Wang (2021) suggest
that when silicon nanoparticles (Si-NPs) are used in moderate amounts, they can
significantly enhance the absorption of nutrients and
promote
fish growth due to their ability to increase feed utilization and nutrient
utilization. Similarly, Liu et al. (2020) experimented to check the
effect of Si-NPs on fish. They confirmed that a medium concentration of Si-NPs
increases feed utilization, nutrient utilization, and growth performance of
fish.
The
study also revealed that higher doses of Si-NPs, such as 3 mg/kg, negatively
impacted the growth performance of GRP4, leading to poorer results. Similarly, Li et al. (2019)
and Chen et al. (2020) Using high amounts of these
nanoparticles has been shown to affect fish growth negatively, because these
nanoparticles can interfere with metabolic processes, slowing them down, and
their toxic nature at higher levels further damages the fish's health. Gao et al. (2021) explored
the effects of sub-lethal levels of Si-NPs on growth-enhancing mechanisms. they
found that Si-NPs positively influenced gut health and enhanced nutrient
absorption. This was evident through the increased bioavailability of essential
amino acids and minerals, which are crucial for growth and development. Liu
et al. (2020) also explained that Si-NPs promote growth, oxidative
stress, and reduction of overall fish health, and they also act as antioxidants.
Zhao et al. (2022) observed that nanoparticles play a role in boosting digestion
and optimizing nutrient absorption. They achieve this by maintaining a healthy
balance of gut bacteria and supporting the proliferation of beneficial
microbial populations. In this study, a
significant reduction in FCR in GRP2 and GRP3 was recorded, indicating the
increased feed efficiency of Si-NPs. Furthermore, the improved FCR indicated
that Si-NPs enhance protein synthesis and significantly increase the
consumption of metabolic energy. On the other hand, metabolic imbalances were
recorded in GRP4 due to increased FCR, indicating that the overload of nanoparticles
makes it less efficient in converting feed to body weight. Hou et al.
(2021) and Zhang et al. (2022) findings indicated that high
concentrations of Si-NPs nanoparticles caused disruption of cell structure,
swelling and can lead to oxidative stress that lessen the growth and survival
and similarly Tacon et al. (2020) also described that changing feed to
body weight more proficiently that indicated the best FCR which is an important
factor in aquaculture profitability.
The current study also described the
effect of Si-NPs on the heavy metals, including both essential and
non-essential trace elements viz; Na, Co, Ni, Fe, Mg, Al, Zn, Pb, Cu, Cr, and
Cd in C. carpio. The findings of the present study are important
in checking the role of Si-NPs in maintaining homeostasis by regulating trace elements,
seeing the ecological toxicology and aquaculture perspectives. Furthermore,
most of the trace elements like Na, Co, Ni, Fe, Mg, Al, Zn, Pb, Cr, and Cd
significantly improved the serum contents in GRP2 (1 mg/kg) and GRP3 (2 mg/kg)
that indicated the moderate levels of Si-NPs increased absorption and
bioavailability of such compounds. Hussain et al. (2024) described that a
medium level of Si-NP nanoparticles improved nutrient absorption in the fish;
similarly, Zhao et al. (2022) also studied the effect of nanoparticles
on heavy metals and indicated increased permeability and absorption of
elements. On theother hand, GRP4 (3 mg/kg) decreases the heavy metals viz; Na,
Ni, Fe, Mg, Al, Zn, Pb, and Cr because due to increased Si-NPs. Torrealba et
al. (2019) and Wang et al. (2024) studied the effect of Si-NPs on
fish. They explained that high concentrations of Si-NPs disturb the homeostasis
of trace elements due to nanoparticle-induced oxidative stress by disruptive
metabolic pathways leading to deficiencies and imbalance. Jan et al. (2015) and Das et al. (2025) also studied the
effect of Si-NPs on heavy metals in fish. They
explained that at high concentrations of Si-NPs, some elements, particularly Fe
and Zn, are significant in antioxidant defence systems and enzymatic functions.
The result also indicated that the effect
of Si-NPs on serum enzyme activities such as (ALT, AST, ACP, ALP, and LDH) revealed significant alterations among the
treatment groups and
the results specify
that moderate doses of Si-NPs reduced tissue damage and inhibited oxidative stress. In the present study,
ALT and AST levels reduced GRP2 and GRP3, indicating that Si-NPs are able to
save liver cells against oxidative damage, thus improving overall liver
function. Similarly, the level of serum LDH activities
was also reduced in GRP2 and GRP3, which may be due to the fact that this
enzyme improved cellular integrity. Farooq and his
colleagues also described that the moderate dose level of Si-NPs on serum
enzymes might be
effective in antioxidant and metabolic activities
(Farooq et al., 2022). On the other hand, the high concentration
of Si-NPs in GRP4 on serum enzymes recorded poor results
that induced harmfulness at higher concentrations. Mehta (2025) observed the drops in the
activities of serum enzymes due to the fact that this nanoparticle has strong
antioxidant activity, scavenging the free radicals and reducing oxidative
stress.
Researchers observed that the high
concentrations of Si-NPs in the fish diet cause enzymes such as ALT and AST to become
the main source of liver damage and are naturally upregulated as a reaction to
oxidative stress (Min et al., 2023). Mahboub et al. (2024) also
studied that Excessive use of nanoparticles can lead to oxidative stress,
inflammation, and damage to cell structures, ultimately inhibiting the health
of tissues.
CONCLUSION:
In the present study, we concluded that
Si-NP dietary supplementation of moderate levels (1–2 mg/kg) significantly
enhanced C. carpio growth performance, feed conversion rate, and
activity of enzymes. Si-NPs also influenced the levels of trace elements, with
enhanced nutrient delivery at optimal dose levels, though causing adverse
impacts at elevated dose levels (3 mg/kg). The findings reveal that Si-NPs can
be a valuable aquaculture feed supplement, but the optimum dosage must be
exercised to prevent potential toxicity. Further investigation is needed to
ascertain long-term safety and impacts on the environment.
Statements And Declarations
Ethical Approval: Department of
Fisheries Saline Water Aquaculture Research Center (APF-024).
Conflict of interest: The authors
declared no potential conflict of interest.
Consent to Participate: All authors have
consented to submit the article to this journal.
Consent to Publish: All authors have
agreed to publish the article in this journal.
Funding: This study did not
receive specific funding from public, commercial, or non-profit organizations.
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