This document reviews the use of nanoparticles as supplements in fish feed to improve growth performance. It discusses how nanoparticles, due to their small size and large surface area, can enhance nutrient absorption and assimilation in fish. Specifically, it examines research on supplementing fish feed with selenium nanoparticles, zinc oxide nanoparticles, and iron oxide nanoparticles. The studies found improved growth metrics like weight gain and feed conversion efficiency in fish receiving nanoparticle-supplemented feeds within optimal dosage ranges. Certain nanoparticles also led to higher nutrient content in fish tissues and improved hematological parameters. In summary, supplementing fish feed with certain metal nanoparticles at the nanoscale appears to boost nutrient bioavailability and fish growth.

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Materials Today: Proceedings 5 (2018) 9076–9081 www.materialstoday.com/proceedings
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Selection and Peer-review under responsibility of 6th
NATIONAL CONFERENCE ON NANOMATERIALS AND
NANOTECHNOLOGY (NCNN VI - 2017 )
NCNN 2017
Nanoparticles as feed supplement on Growth behaviour of
Cultured Catfish (Clarias gariepinus) fingerlings
Onuegbu Chris U1*
., N.B.Singh1
and Anupam Agarwal2
1
Research and Technology Development Centre, Sharda University, Greater Noida, India.
2
Chemistry Department, SBSR, Sharda University greater Noida, India
Abstract
The health and economic importance of catfish and its advantages as a cultivable species have been discussed. The major
challenge facing catfish farming is availability of relatively cheap but high quality feed. Traditionally, feeding fish has relied on
providing fish with food in the form of a food pellet. This pellet is chiefly formulated based on the daily nutritional fish
requirements for components such as fats, proteins, carbohydrates, minerals and vitamins. The applications of nanomaterials as
supplements in fish feeds, and their effects on fish body chemistry have been reviewed. Nano-sized (10−9
–10−7
m) particles offer
many technical and biomedical advances over the bulk material. They have unique properties because of their small size and
large specific surface area. Fish farming is acknowledged as an important and substantial part of the global fishing industry. It is
found that nanoparticles will enhance aqua feeds by increasing the proportion of fish food nutrients that pass across the gut tissue
and into the fish, rather than passing directly through the fish digestive system unused. Dietary minerals at the nanoscale size
may pass into cells more readily than their larger counterparts. This accelerates their assimilation process into the fish. Adding
mineral nutrients to the regular fish diet at the nanoscale might also have a tremendous impact not only on growth but also on the
overall health of the fish. Number of nanomaterials as supplement for fish feed on growth performance have been discussed.
© 2017 Elsevier Ltd. All rights reserved.
Selection and Peer-review under responsibility of 6th
NATIONAL CONFERENCE ON NANOMATERIALS AND
NANOTECHNOLOGY (NCNN VI – 2017)
Keywords: 8pt Aquaculture; Nanoparticles; Catfish; Nutrition; Growth Performance
*Email address: ocaesars47@gmail.com

2. Onuegbu Chris U / Materials Today: Proceedings 5 (2018) 9076–9081 9077
1.0 Introduction
Fish is a very important source of relatively cheap but high quality protein especially for poor people [1]. For some
time, now fish supplies from natural marine and freshwater sources (capture fisheries) have been on the decline due
to sharp increase in demand because of rise in per capita consumption [2]. This has necessitated artificial cultivation
of fish called as aquaculture. The biggest challenge that threatens sustainable aquaculture development is the
availability and affordability of high quality feed. The basic nutrient requirements of fish include protein (32%),
dietary energy (8.5-9.5%), fat (4-6%), carbohydrate (20-35%) and fibre (< 4%), but vary in accordance with age and
species [3]. Several combinations of plant and animal ingredients have been used to have optimal nutrient
requirements in fishes. The ideal fish feed is formulated with fishmeal; but due to its cost and declining fish
supplies, combinations of different plant and animal products have been used to improve nutrient (particularly
protein) content in fish feed. Many feed ingredients contain some essential fish nutrients in desired amounts but the
nutrients have low bioavailability in fish [4]. Consequently, minute concentrates of some soluble organic/inorganic
materials have been used as additives in fish feed either to augment deficient nutrients or act as catalyst to improve
nutrient bioavailability. Recent advances in nanotechnology are currently revolutionizing different aspects of
science and technology, including aquaculture.
The fisheries and aquaculture industries can be revolutionized by using nanotechnology with new tools, such as
rapid diagnosis of diseases which will enhance the ability of cultivable organisms to uptake drugs like hormones,
vaccines and essential nutrients etc., [5]. The metal nanoparticles (NPs), such as Fe, FeO, Se, Zn, ZnO, Cu, and
MgO etc., play a crucial role in aquaculture operations. It has been reported that iron NPs when fed to young carp,
Carassius auratus and sturgeon, Acipenser gueldenstaedtii showed a faster growth rate, 30% and 24% respectively
[5]. Different Se sources (nano-Se and Selenomethionine) supplemented diets improved the growth, antioxidant
status and muscle Se concentration of Crucian carp, Carassius auratus [5]. In recent reports, dietary
supplementation of Zn, nano Zn and Cu have produced better survival and growth in M. rosenbergii PL [5].
Nanotechnology is in constant development and its applications are ever more varied and specific, with a high
potential for improving livestock production and animals in general. The study of nanotechnology in these areas is
still very limited. In this article, the recent research on the effect of nanomaterials as feed supplements to fish
(particularly catfish) has been reviewed.
2.0 Fish feeds
Fish are unable to synthesize their food. They depend on the food chain for their food needs. Fish are omnivorous
feeders and, in their natural habitats, can ingest a wide range of plant and animal materials. In culture, however,
deliberate efforts are made to enrich fish feeds with essential nutrients for faster and healthier growth and better
returns. Under semi-intensive culture, fish may be fed supplementary feeds which consist of cheap and locally
available materials like kitchen wastes, agricultural products etc. to augment what is naturally available. Under
intensive culture, however, they are fed complete feeds which are a mixture of carefully selected ingredients that
provide them with all nutrients essential for their fast and healthy growth. Fishmeal is considered an ideal source of
dietary protein for fish but sustainability of its use is challenged by dwindling capture fisheries resources and high
costs [6,7]. The relative percentage of the nutrients in fish feed depends on the age, sex and species of fish involved.
Reports have shown that different conventional feeds formulated with different ingredients or different proportions
of same ingredients result to different growth performances and body compositions of fish [8-10]. Table 1 shows the
proximate composition of four typical complete feeds [11].
Table 1: Proximatecompositions of the four commercial feeds [11].
Parameter T1 T2 T3 T4
Moisture content (%) 8.53 8.07 8.67 8.67
Protein (%) 46.36 45.64 48.10 47.67
Ether extract (%) 28.50 29.63 30.37 27.73
Ash (%) 9.38 8.53 8.67 7.77
Crude fibre (%) 7.23 8.13 6.83 7.57
Energy (Cal/kg) 209.24.23 190.12 250.67 230.45

3. 9078 Onuegbu Chris U / Materials Today: Proceedings 5 (2018) 9076–9081
3.0 Nanomaterials
These are materials having one or more of their dimensions in the nanometre (1-100 nm) range. In nano-size range,
materials have been associated with novel properties and applications which are different from their bulk
counterparts. Because of their small size, nanomaterials possess very high exposed surface area per unit volume
which enhances their chemical reactivity. They are also reported to be very stable under conditions of high
temperature and pressure. These are easily taken up by the gastrointestinal tracts in animals and so are more
effective than bulk materials at lower doses [11], interact more effectively with organic and inorganic materials in
animal bodies [12] and can cross the small intestine and further distribute into the blood, brain, liver and other
organs of the animal body [13]. Nanomaterials for use in animal feeding are preferably prepared by chemical or
biological methods for particle size uniformity and environmental friendliness respectively [14]. However, chemical
method may involve toxic and/or corrosive substances while prolonged particle formation time, culture
condition/media maintenance and problems of products recovery from solution are some of the challenges of the
biological methods [15].
4.0 Fish Growth
Growth in animals may be regarded as an improvement either in size (weight, length and girth) or in health (body
biochemical constitution harmony) or both. A sound understanding of both the concept and importance of growth is
central to meaningful improvement/progress in fish nutrition. Growth is important in the estimation of production,
stock size, recruitment and mortality of fish populations in fisheries and aquaculture. Several indices have been
evolved for the expression of physical growth in fish. These include weight gain (WG), percent weight gain (%
WG), average daily gain (ADG), specific growth rate (SGR), feed conversion ratio (FCR), protein efficiency (PE)
and protein production value (PPV) [16,17]. On the other hand, the health status of fish is monitored and expressed
using biochemical indices such as proximate body compositions, mineral accumulation in certain vital organs or
tissues, haematological and immunological parameters among others. A number of nanomaterials are currently
being employed as feed supplements and are discussed under the following heads.
5.0 Selenium nanoparticles
Selenium is an essential micronutrient which enhances the growth and physiological health of fish by acting as an
antioxidant as well as a catalyst for growth and hormone production [18]. SEM and TEM of selenium nanoparticles
are given in Fig.1 [19].
(a) (b)
Fig.1 (a) SEM and (b) TEM images of selenium nanoparticles [19].
Recently, the synergic effect of dietary nano-selenium (nano-Se) and vitamin C on the growth of mahseer fish (Tor
putitora) has been studied [20]. Reports showed that percent weight gain (%WG), feed conversion efficiency (FCE
%), and specific growth rate (SGR) of fish were significantly improved (p < 0.05) with nano-Se supplementation in
feed up to the level of 0.68 mg n-Se/kg dry feed, compared to fish feed basal diet; but feed conversion ratio (FCR)

4. Onuegbu Chris U / Materials Today: Proceedings 5 (2018) 9076–9081 9079
was significantly lower (p < 0.05) in fish fed supplemented diet against those fed basal diet. Also study conducted
on common carp (Cyprinus carpio) with different levels of nano-Selenium reportedly [21] showed improved growth
performance (in terms of final weight and weight gain) as well as higher selenium contents in liver and muscle in
fish fed diet supplemented with nano-Selenium at 1 mgkg-1
dry feed level compared to the control. Significantly
higher (p < 0.05) total protein and globulin but lower albumin were equally reported for supplemental level of 2 mg
kg-1
dry feed. Antioxidant activities were generally significantly (p < 0.05) improved in fish fed 1 and 2 mgkg-1
dry
feed compared to diets containing higher or lower levels of nano-Se supplementation.
6.0 Zinc
Dietary zinc is very essential for growth and health of higher animals, including fish. Zinc promotes growth, acts as
antimicrobial agent and modulates both immune and reproductive systems of animals. Zn cannot be stored in animal
body and its deficiency could lead to frequent infections, poor appetite as well as taste and smell associated
problems. Regular dietary zinc intake is therefore a necessity [22,23]. The effects of nano zinc oxide (nZnO) on the
growth and haematological parameters of grass carp (Ctepharyngodon idella), have been investigated against ZnO
and ZnSO4, as dietary zinc supplements in basal feed [24]. Two levels of supplementation (30(level 1) and 60(level
2) mg Kg-1
) were considered for each treatment. Percent weight gain (%WG), specific growth rate (SGR), and feed
conversion ratio (FCR) were reported to be significantly (p < 0.05) higher in fish fed level 1, followed by level 2 of
nZnO supplemented diets. Results have shown that growth was retarded in fish fed both levels of ZnSO4 and level 2
of ZnO. Again supplementations of ZnSO4 and ZnO at both levels and nZnO level 2 significantly decreased
haematological parameterssuch as red blood cells (RBCs) and white blood cells (WBCs). This suggests 30 mg/Kg
feed to be the optimal dietary supplementation level of nZnO for C. idella. An X-ray diffraction pattern of ZnO
nanoparticles is given in Fig. 2 [25,26]
(a) (b)
Fig. 2: (a) SEM image and (b) XRD pattern of ZnO NPs [25,26]
7.0 Iron
Iron is important in the life of fish because of its role in physiological processes of oxygen transport, cell respiration,
lipid oxidation reactions, immune system functioning and defence against infections [27-29]. Dietary iron
supplementation for fish is essential because dietary requirements are not met by most natural bulk iron sources due
to low solubility and low bioavailability [29]. The growth promoting potentials of iron oxide NPs (nFe2O3) in
freshwater prawn (Macrobranchium rosenbergii) post larvae have been studied against a basal diet [30]. The result
showed significant improvement (p < 0.05) in survival rate, growth, digestive enzyme activities, body biochemical
constitution and some haematological parameters of the fish fed supplemental levels 10-20 mg nFe2O3 Kg-1
dry feed
compared to those fed with basal feed. Negative responses were however reportedly obtained at higher supplemental
levels, 30-50 mg nFe2O3 Kg-1
dry feed. Earlier, [31] compared the performance of Indian major carp (Labeo rohita)
under a basal feed and two diets (basal feed supplemented with 0.54 mg nFe2O3 Kg-1
feed and 0.55 mg FeSO.7H2O

5. 9080 Onuegbu Chris U / Materials Today: Proceedings 5 (2018) 9076–9081
Kg-1
feed respectively). Figure 1 and 2 show the growth performance and haematological profile of Labeo rohita H
fed nFe2O3 and FeSO.7H2O supplemented diets as T1 and T2 respectively (Tables 2 and 3). They reported significant
improvement in survivability, growth and haematological parameters of fish fed nano-iron supplemented diet
compared to the control but no significant difference between the performances of fish fed diets containing 0.54 mg
nFe2O3 Kg-1
feed and 0.55 mg FeSO.7H2O Kg-1
feed. Some haematological parameters (RBC, WBC, Hb and HCT)
of freshwater fish, Oreochromis niloticus were reported to have been significantly negatively when they were
exposed to biologically synthesized α – Fe2O3 NPs at concentrations levels of 0.5, 5 and 10 µg/ml [32]. Blood
parameters in Indian major carp (Labeo rohita) were differently affected when the fish was exposed for 25 days to
environment (water) containing 500 mg of nFe2O3/litre of water.
Table 2: Growth performance and survivability of Labeo rohita
Supplemented with different iron sources (T1 and T2) [31]
Group/treatment T1 T2 Control
Initial weight 20.22 ± 0.44a
20.29 ± 0.68a
20.45 ± 0.50a
Final weight 38.07 ± 0.90a
36.00 ± 0.00a
28.25 ± 0.88b
Survival rate 100a
100a
100a
Table 3. Haematological parameters and total serum proteins of Labeo rohita H. supplemented
with different iron sources (T1 and T2) and without iron (control) in the basal diet [31]
RBC (106
)
cells/µl
WBC (106
)
cells/µl
Haemoglobin
(g/dl)
Total protein
(g %)
Albumin
(g %)
Globulin
(g %)
Control 1.60 ±0.15a
102.38±0.72a
7.13 ± 0.10a
3.61±0.02a
1.23 ±0.01a
2.38 ±0.02a
T1 2.92±0.02b
102.50 ± 0.16a
11.53 ± 0.11b
4.70 ± 0.00b
1.12 ± 0.00b
3.59 ± 0.00b
T2 2.61 ±0.02c
100.45 ±1.55a
10.40 ± 0.16c
4.63 ± 0.01b
1.15 ± 0.00b
3.49 ± 0.02b
Superscripts a and b were significantly different (p < 0.05)
White blood cells (WBCs) counts were higher but red blood cells (RBCs) counts lower in fish in the control
compared to these in the treated water throughout the period [33]. Mean corpuscular haemoglobin concentration
(MCHC) values remained fairly the same and constant during the trial period in both the treatment and control.
Other blood parameters, mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and haematocrit
(Hct) were higher for the first couple of days in the treatment but declined thereafter in favour of the control.
Closely related results were reported by [34] who also exposed Indian major carp (Labeo rohita) to different
concentrations (1 and 25 mg/l) of nFe2O3 for 96 h. Different haematological parameters were differently by the
control and the two treatments.
8.0 Conclusions
Mineral nanoparticles are proving more effective than their bulk counterparts as feed supplements, in improving the
growth and health of cultured fish. This higher level of effectiveness is interestingly observed with lower doses of
nanomaterials which make them even more cost and material effective than the bulk materials. Higher doses of
nanomaterials were found to be detrimental to fish growth and health. Nanotechnology is a recent way. Different
species and ages of fish were found to be differently impacted by nanomaterials of same nature, source and particle
size. Conversely, same species and age of fish were affected in different ways by different ways by nanomaterials of
different natures, sources and particle sizes. Hence for sustainable improvement in the growth and health of a
particular fish by nanomaterial supplementation in feed, the particle size and dose of the nanomaterial is to be
worked out.
Despite their acclaimed universal spread, high cultivable attributes and global health/economic importance, there are
yet no available records of scientific studies of the possible effects of nanomaterials, as feed additives, on the growth
and health of the catfish. This is therefore a virgin area of research for aquaculture nutritionists and scientists.

6. Onuegbu Chris U / Materials Today: Proceedings 5 (2018) 9076–9081 9081
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