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May | June 2014

Prospects on dietary trace minerals:
Aquafeeds & Aquaculture

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FEATURE

Prospects on dietary trace minerals:

Aquafeeds & Aquaculture
by Sungchul Charles Bai, Professor/Director, Deptartment of Marine Bio Materials & Aquaculture, Feeds and Foods Nutritional
Research Center (FFNRC), Pukyong National University, Nam-gu, Busan, Korea

Dietary trace minerals must be supplied in adequate quantity to ensure the optimal
growth and health of farmed aquatic species. The significance of trace mineral
bioavailability has become more important as the aquaculture practice has gone under
phenomenal intensification worldwide. Moreover, the composition of majority of
commercial fish feed has been changing and there is an increased use of dietary plant
protein. As a result, the bio-availability of trace minerals are being adversely affected
by the presence of antagonistic factor such as phytic acid in plant protein.

ven though, our knowledge in fish

nutrition has advanced significantly,
the information on trace minerals
requirement is still limited and
fragmentary.
Whereas, the sustainability issue has put
a new dimension in aquafeed formulation
with a wide array of new ingredients and
additives, on the other hand, the importance
of basic nutrient such as trace minerals is still
in sideline.
Substantial investment and integrated scientific efforts are warranted to bridge the
knowledge gap and further improve our
understandings on the significance of dietary

trace mineral in fish nutrition and health at the

least cost to environment.

An overview

The importance of trace minerals supplementation in fish feed formulation has

been well accepted since some of the trace
minerals from ambient water and feed itself
cannot supply the optimal level required by
the cultured aquatic species.
The reluctance among researchers to
determine the trace mineral requirement has
been partly due to the related difficulty of
conducting research on mineral nutrition.
Problems associated with the quantification
of mineral requirements include identification

Table 1: Trace Minerals their functions and deficiency signs in aquatic species
Microminerals/
Trace Minerals

Functions

Deficiency Signs

Impaired growth and reduced

activity of copper-containing
enzymes

Copper

Metalloenzymes,
Fe metabolism

Cobalt

Vitamin cyanocobalamin ( B12 )synthesis

Chromium

Carbohydrate and lipid metabolism

Anemia
Impaired glucose utilization

Iodine

Thyroid hormones, energy production

Thyroid hyperplasia

Iron

Hemoglobin, enzyme, lipid oxidation

Impaired growth, anemia

Manganese

Organic matrix of bone

Impaired growth, skeletal

abnormalities, cataracts

Molybdenum

Xanthine oxidase

Reduced enzyme activity

Selenium
Zinc

Glutathione peroxidase, component of

amino acid selenocysteine
Metalloenzymes,

Reduced growth, anemia,

exudative diathesis, reduced activity
of glutathione peroxidase
Reduced growth, anorexia,
cataracts, skeletal abnormalities,

Source: Adopted and Modified from NRC, 2011 and S. C. Bai, 2011, Nutrient Metabolism &
Feeds in Fish, pp. 41
18 | INTERNATIONAL AQUAFEED | May-June 2014

of the potential contribution of minerals from

the water, leaching of mineral from the diet
prior to consumption, availability of suitable
test diets that have a low concentration of
the targeted mineral and limited bioavailability
(NRC, 2011).
The trace minerals including copper, manganese, iron, zinc, selenium, chromium, iodine
and fluorine participate in a variety of metabolic processes. Some of the vital biochemical
processes involving minerals are the formation
of skeletal structures and other hard tissues
(ex. Fin, rays, scales, teeth and exoskeleton),
electron transfer, regulation of acid: base
equilibirium, the production of membrane
potentials and osmoregulations.
In strict sense, except the osmoregulation,
rest of the basic metabolic functions of various
elements are same for aquatic and terrestrial
animals. Trace minerals or micro-minerals are
important components of hormones and
enzymes serve as cofactors and/or activators of a variety of enzymes as well as
participate in a wide variety of biochemical
processes (NRC, 2011). Only a few scientific review including, Davis & Gatlin (1996)
and NRC 2011 summarizes the estimate of
mineral requirement in few commercially
important aquaculture species.
However, the trace mineral requirements in several other aquaculture species
are still unknown and the practical feed
formulation follow the general estimation.
Table 1 summarises the function and
common deficiency symptoms of trace
minerals in aquaculture. The deficiency
symptoms depend upon the degree and
duration of deprecation of that certain trace
mineral in certain species under certain
environmental condition.
In strict sense, one cannot define the
sub-optimal level of trace mineral for an
aquaculture species applicable for all kind
of culture environment. The requirement

FEATURE
level and deficiency symptoms for
trace minerals under appropriate
and stressful environmental condition remains to be investigated
in aquaculture.
At this point, we must appreciate what is known today and
try to further improve our understanding in the significance of
trace mineral in fish nutrition,
health and environment.

Lower requirement

Even though, trace minerals

are typically required in much
lower quantity in fish diet but
its supplementation at optimum
level is a critical issue in fish feed
formulation.
Scientific reports gathered
over last two decades indicate,
few trace mineral are quite sensitive and proper care should be
taken to ensure their inclusion
at optimum level in fish feed
formulation. For instance, trace
mineral selenium (Se) is an essential micronutrient in animals and is
required for normal growth and
development.
However, high selenium concentrations in an animals diet can
result in toxic effects.
Selenium is similar to sulfur
with regard to its basic chemical
and physical properties (it has
the same valence states, forms,
and analogs of hydrogen sulfide,
thiosulfate, sulfite, and sulfate)
and mammalian studies show
that cells do not discriminate
well between the two elements
as proteins are being synthesized
(it is assumed that the mechanistic features underlying toxicity
are essentially the same for fish,
because the resulting pathology
and teratogenic features are the
same).
When present in excessive
amounts, selenium is erroneously
substituted for sulfur, resulting in the formation of a triselenium linkage (Se-Se-Se) or a
selenotrisulfide linkage (S-Se-S).
Either configuration prevents
the formation of the necessary
disulfide chemical bonds (S-S).
The result is distorted, dysfunctional enzymes and protein
molecules, which impair normal
cellular biochemistry (Ganther
1974; Stadtman 1974; Diplock
and Hoekstra 1976; Reddy and
Massaro 1983; Sunde 1984).

Consequently, there has been

public awareness and governmental efforts including in the
Republic of Korea to establish
an upper limit of selenium in
aquafeeds to prevent both cultured fish and consumers from
selenium toxicity.
Consecutive studies conducted in my laboratory, investigated
the requirement and toxicity
level of dietary Se in few commercially important species (Kim
et. al., 2003; Lee et al., 2008; Lee
et. al., 2010; Arshad et. al., 2010).
Based on our experimental
results, we concluded that a
dietary Se level above 7.38mg
Se/kg is likely toxic and with a
long-term feeding trial, a dietary
Se level of 4.13mgSe/kg may
cause toxic effects in juvenile
olive flounder.
While the overall performance from our preliminary
feeding trail, a dietary Se level
of 0.21mg NaSeO3/kg diet was
concluded to be optimal for
proper growth performance and
a dietary Se level of 12.3mg Na2
SeO3/kg was anticipated to be
toxic to juvenile black seabream.
Likewise, various other studies have reported the toxicity
level for Se in different aquaculture species. Overall observations in our experiments in
different species and taking into
account other reports, it appears
the requirement and toxicity
level for Se is species specific and
therefore proper care should be
taken to avoid its adverse effects.
Further few other trace
mineral such as dietary Copper
(Cu) toxicity has also been documented by various other authors.
Therefore, dietary trace minerals
should be supplemented in judicious manner, keeping in mind a
delicate balance must be maintained between the ingestion,
digestion and absorption of trace
minerals.

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Fish health management

The concept of maintaining the health of fish through

the best possible nutrition is
well-accepted in modern fish
farming.
Scientific evidence gathered
over the past 30 years indicates
that dietary nutrients as well as
additives could stimulate the

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May-June 2014 | INTERNATIONAL AQUAFEED | 19

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3/18/14 4:33 PM

FEATURE
Table 2. Growth performance and hematological characterstics of juvenile Black seabream fed
different levels of dietary Se for 15 weeks.
Diets1

WG(%)

FE(%)

SGR(%)

PER

PCV(%)

Hb(g
/100ml)

RBC
(X106cell/
l)

Se 0.21

327.4a

93.3a

2.70a

1.72a

42.2

15.3

3.34

Se 0.30

357.5a

94.9a

2.88a

1.79a

38.2

16.5

3.69

Se 0.52

325.6a

91.9a

2.69a

1.72a

38.6

16.1

3.59

Se 1.29

349.7a

94.3a

2.80a

1.78a

32.5

17

3.12

Se 12.3

254.4b

84.2b

2.23b

1.53b

26.6

13.1

2.9

13.0

1.40

0.08

0.03

2.46

0.72

0.16

Pooled SEM9

1Diets Se 0.21, Se 0.30, Se 0.52, Se 1.29, Se 12.3 contained 0.21, 0.30, 0.52, 1.29, and 12.3
mg sodium selenite (Na2Seo3)/kg diet.
Note: For more detail, please refer Lee. et. al., 2008

immune system of fish and help to fend off

diseases (Kiron, 2014).
Consequently, the last decade has witnessed a great deal of attention paid to develop safe and cost effective immunostimulants,
probiotics, prebiotics, synbiotics and so on.
Worthy to note that functional feed has
been defined as the feed which can supply the
nutrient beyond the basic requirements of a
cultured species. Nutritionally well-balanced
and properly processed diets are the prime
importance in intensive aquaculture, where
fishes are frequently reared in stressful environment.
Deficiency of any nutrients, especially vitamins and micro minerals could be an ample
reason to trigger the disease epidemic in an
aquaculture operation.
Trace minerals being the integral part
of several metalloenzymes have significantly
greater impacts on preventive health management and success of an aquaculture venture.
For example, Lim et al., (2001a) attributed
the imbalances in Iron (Fe) would compromise the immune system and the resistance
of fish to disease. The effects of dietary zinc
on immune response and disease resistance
in fish has also been reviewed by Lim et al.
(2001b).
Observations reported include enhanced
chemotaxis of macrophages, a lower phagocytic ability, improved or attenuated disease
resistance and reduced or negligible effect on
antibody production.
Selenium is another important trace element for fish because it is a constituent of
selenoproteins and has structural and enzymatic roles similar to glutathione peroxidase
(the antioxidant enzyme). This mineral modulates the immune functions such as inflammation and virulence development (Rayman,
2000; Kiron, 2012).
In channel catfish selenoyeast and selenomethionine as the source of Se were
observed to increase the antibody titer corresponded to their dietary concentrations
(Wang et al., 1997).
Furthermore, Se is an integral part of the

enzyme glutathione peroxidase (GSH-Px),

and it has some complementary biochemical
functions with vitamin E (Gatlin et. al., 1986a).
GSH-Px is part of the cellular defence system
against oxidative damage, together with the
antioxidant vitamins such as vitamins C and E.
Hilton (1989) mentioned that these nutrients
do not act independently and are interrelated
with other nutrients in terms of function and
metabolism.
A few studies on the interactions of these
vitamins in fish have been investigated in
rainbow trout, Salmo gairdneri (Richardson)
(Hung & Slinger 1980) and channel cat fish,
Ictalurus punctatus (Rafinesque) (Gatlin et.
al.,2003 1986b).
Vitamin E and Se function synergistically in
animal tissues to form an important antioxidant defence system. The interactions of Se
and vitamin E in fish have been reported in
channel cat fish (Gatlin III et al. 1986a; Wise,
et. al., 1993), Atlantic salmon, Salmo salar L.
(Poston et. al., 1976), rainbow trout (Bell et.
al., 1985) and chinook salmon, Oncorhynchus
tshawytscha (Thorarinsson et. al., 1994).
In addition, high dietary supplementation

of vitamins C and/or E and Se showed positive effects on growth and immune response
in brook trout, Salvelinus fontinalis (Mitchill)
(Poston & Livingston1969), channel cat fish
(Durve & Lovell1982; Li & Lovell 1985; Li
et. al., 1993), Atlantic salmon (Salte et. al.,
1988; Hardie et. al., 1990) and rainbow trout
(Navarre & Halver 1989).
In a study conducted in my laboratory
to study the synergetic effects of vitamin C,
E and selenium, observation suggested that
dietary supplementation of vitamin C and E
over required values had positive effects on
growth performance, but there were no synergetic effects of dietary vitamin C, E and Se
supplementation over minimum requirement
levels on the growth performance and disease
resistance in fingerlings Nile tilapia.
Altogether scientific evidences suggest, in
depth investigations are warranted to explore
the opportunities to use the dietary trace
mineral in preventive health management in
aquaculture.

Plant protein in aquafeed & trace

minerals bioavailability

Feed formulations for farmed aquatic animals have historically relied on fishmeal to
provide a major part of their nutrient requirements.
However, economic and sustainability
issues have exerted substantial pressure for
the reduction of fishmeal in aquafeeds.
Numerous scientific studies in last three
decades have investigated the efficacy of
different plant protein as an alternative to
fishmeal.
Consequently, there has been a massive
shift towards the use of plant protein in
aquafeed formulation. Plant protein contain a
wide array of antinutrients, among them the
presence of antagonists factors such as phytic
acid has been acknowledges as the major

Table 3 . Efficacy of chelated trace minerals in aquaculture

Minerals

Fish/Crustacean

Remarks1

Reference

Se

Channel catfish

Higher Bioavailability

Wang & Lovell, 1996

Zn

Channel catfish

Equivalent

M.H.Li & Robinson , 1996

Zn

Abalone

2~3 times

Tan & Mai, 2001

Cu

Grouper

2~4 times

Lin et al., 2010

Cu

Olive flounder

2 times

Mohseni et. al., 2011

Cu

Sturgeon

2 times

Mohseni et. al., 2011

Cu

Pacific white
shrimp

3~4 times

Bharadwaz et al., 2014

Korean rockfish

2~4 times

Katya et. al.,2014

Premix (Cu, Zn
& Mn)

Pacific White
shrimp

4~6 times

Katya et. al., 2014 (Unpublished)

Premix (Cu, Zn
& Mn)

Rainbow trout

2 times

M..J.S. Apines et al., 2003, 2004

Premix (Cu,
Zn, Mn & Fe)

1 Remarks represent the reported comparative efficiency of chelated trace mineral Vs Inorganic
source in respective experiment
20 | INTERNATIONAL AQUAFEED | May-June 2014

FEATURE

barrier hindering the bioavailability of trace lower Ph in fish gut apart from its high cost.
mineral. Phytic acid (myo-inositol 1,2,3,4,5,6- Deterioration of phytase activity at high temhexakisphosphate) is the major phosphorus perature of fish feed processing, especially in
(P) storage compound in plant seeds and can the case of extrusion processing are additional
account for up to 80 percent of total phos- factors, all together limiting the use of phytase
phorus. Phytic acid binds with divalent cationic in aquafeed formulation.
Another approach to increase the bioavailtrace minerals rendering them unavailable to
the animal and these are consequently lost to ability of trace mineral as highlighted in NRC
the environment as waste (Cheryan, 1980; (2011), as the aquatic animal feed industry
Davis and Gatlin, 1996; Davis et al., 1993; Li increase its use of plant feedstuffs, the need
for mineral supplementation should increase.
and Robinson, 1997).
Since environmental pollution due to high
The limited bioavilability and potential
dietary deficiency of trace minerals are serious rate of mineral excretion by mineral antagoconcern as adequate trace mineral ensuring nisms at higher level of dietary inclusion
the optimum growth and health of cultured has been a common problem in livestock
husbandry. It remains an important research
species in aquaculture.
An ideal approach to improve the bio- area for scientific community to clearly underavailability of trace mineral has been recom- stand the ultimate fate of trace minerals at
mended as the inclusion of microbial phytase higher level of dietary inclusion in aquafeed
in fish diet. Phytase is an enzyme chemically formulation.
known as myo-inositol-hexaphosphate phosphohydrolase (Class 3: Hydrolases), produced Trace mineral form and sources:
either by microorganisms or present in some Critical issue
plant ingredients.
Inorganic form (sulfate/nitrate) of trace
Monogastric animals cannot produce this mineral has traditionally been used in aquaenzyme.
feed formulation.
Presence of phytase in Hatchery Ad Campaign | Theme:limited Shellfish Diet
some animals is of
However, the Ad-1 bioavailability of
Reed Mariculture, Inc. | 2014
microbial origin. Microbial phytase either as inorganic source of trace mineral due to its
lacement: International Aquafeed | Size: Half Page | Dimensions: 190mm X 132mm
a dry powder or as a liquid is available com- higher affinity to antinutrients has hastened
mercially ( Baruah. et. al., 2004).
the search for alternative form of inorganic
However, microbial phytase has been trace minerals.
reported to have limited efficiency due to
As a result, scientific communities have

attempted to develop more stable and bioavailable form of trace minerals suitable for
aquaculture. For instance, tri-basic copper
chloride (TBCC) has been reported as the
more concentrated form of copper than copper sulfate (58% vs 25% Cu).
Since it has low hygroscopicity and is
insoluble in neutral water, it should be a less
reactive and less destructive form of copper when combined with vitamins in diets
(Cromwell et al., 1998). Shao et. al., (2010)
suggested TBCC could be a new dietary copper source as more bioavailable than copper
sulfate for crucian carp.
However, the supporting information is
scanty and needs further research to justify
the bioavailability of TBCC in aquaculture.
In last decade, research is increasing shedding light on the potential benefit of using
organic/chelated form of trace mineral in
aquaculture.
Typically, organic trace minerals are more
stable in the digestive tract and less prone
to interactions and antagonisms as they are
bound to organic molecules and less available to interaction and binding. Some of
the commonly Version: 1
Design: A | available organic trace minerals are metal proteinates, metal amino acid
complexes and metal amino acid chelates.
Earlier studies have demonstrated improved
bioavailability, growth and disease resistance
in fish fed metal proteinates (zinc protein-

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FEATURE

EXCELLENCE IN YEAST
EXCELLENT FOR FISH
Figure 1: Average mortality (Mean SD) of juvenile olive
flounder exposed to dietary selenium for 10 weeks.
Diets Se0.61, Se4.13, Se7.38, Se18.6, Se35.9, Se66.0, and
Se146, contained 0.61, 4.13, 7.38, 18.6, 35.9, 66.0, and146 mg
Se/kg diet respectively
Note: For more detail, please refer Lee. et. al., 2010

Figure.3. Survival rate (%) of Olive flounder fed different

levels of dietary chelated Cu for 12 weeks.

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(CuM20), 43.4 (CuM40), 82.1 (CuM80), 158 (CuM160), 308
(CuM320), 658 (CuM640),
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Aquakultur 90 x 270 AquaFeed magazine.indd 1

ate) and metal amino acid

complexes (zinc methionine)
compared to fish fed inorganic
sources (Hardy and Shearer,
1985; Paripatananont and Lovell,
1995a,b, 1997).
The glycine chelates of trace
minerals have been shown to
improve performance, tissue
mineral retention, hematology
parameters, immune function
and disease resistance in the
rainbow trout (Apines et al.,
2003; Apines-Amar et al.,
2004a,b; Satoh et al., 2001)
and in red sea bream (Sarker
et al., 2005), despite the presence of dietary antagonists
such as phytic acid or tricalcium phosphate (Bharadwaz
et. al., 2014).
Table 3 summarises the few
scientific reports on the efficacy
of chelated mineral in aquaculture. Nevertheless, a cross comparison among these reports on
chelated trace minerals could be
perhaps misleading.
Since, fish species, fish size,
diet composition and other
experimental condition vary
among different experiments.
Further, a fundamental difference among different experiment comparing chelated trace
mineral and inorganic trace minerals is the balancing of ligand in
all experimental diets has been

22 | INTERNATIONAL AQUAFEED | May-June 2014

10.01.14 10:03

overlooked by various research

groups.
Since, ligand could be an
additional source of nutrients
in chelated trace minerals,
researchers must give importance to balancing the ligand in
inorganic treatment as well, to
prevent the bias that difference
in results could be due to ligand.
Available scientific reports particularly published in last decade
evidenced that there is a significant increase in the efficiency of
chelated minerals in aquaculture.
Nevertheless, every manufacture claim their product to be
best and making a right choice
is often a challenge. Recently,
consecutive studies in my laboratory investigated the efficacy
of chelated minerals consisted
of a range of divalent cationic
minerals chelated to two molecules of HMTBa (2-hydroxy4-methylthiobutanoic acid or
hydroxy analog of methionine;
Mintrex, Novus International,
St. Louis, USA) claimed to have
in an extremely stable configuration.
The stability of these molecules renders chelated trace
minerals less available to binding
to phytic acid and to interference from other dietary antagonists.
These molecules are thus

FEATURE

Figure 2: Cumulative mortality rate (%) of Nile tilapia

challenged by E. tarda.
Diet Control: (150mg AA,100mgTAand 0.2mg Se per kg diet)
Excessive ascorbic acid (eAA): (2000 mg AA, 100mg TA and 0.2mg Se per kg diet)
Excessive a-tocopheryl acetate (eTA): (150mg AA, 240mg TA and
0.2mg Se per kg diet) Excessive selenium (eSe): (150 mg AA, 100 mg
TA and 0.5 mg Se per kg diet)
Excessive all (eALL): (2000 mg AA, 240 mg TA and 0.5 mg Se per kg diet)
Note: for more detail please refer, Kim et. al., 2003

able to reach the receptors in the

gut epithelium where they are
absorbed into the circulation of
the animal (Eide, 2004; Wang and
Zhou, 2010; Yi et al., 2007).
Overall performance observed
in our experiments vouched the
potential benefit of using chelated trace mineral, Mintrex Cu
in marine fish, Olive flounder and
fresh water fish white sturgeon
(Mousheni et. al., 2011), Mintrex
Cu, Zn, Mn & Fe premix in Marine
fish Korean rockfish (Katya et. al.,
2014) and Mintrex Cu, Zn & Mn
premix in marine shrimp, Pacific
white shrimp (unpublished).
Worthy to note that, chelated
trace minerals and their premix
should also be supplemented at
optimum level, high level of dietary inclusion could also be toxic.
Table. 4. shows the toxic
effects of chelated mineral premix observed in Olive flounder
(Mohseni et. al., 2012).
Overall trend shows, chelated
trace minerals and premix is holding a great potential as safe and
effective alternative to traditional
inorganic source of trace minerals in aquaculture. Even though
despite of potential benefit, the
high cost of chelated trace mineral often acknowledged as the
major factor limiting its use in
aquafeed industry.
Thus, complementary inclu-

sion of chelated with inorganic

trace minerals could be logical
step to encourage the inclusion
of alternative dietary mineral
source in aquafeed formulation.

Conclusion

The significance of dietary

trace minerals should get due
recognition, for the vision of
aquaculture sustainability.
The true value of any high
quality fish feed is non-existent,
if the basic nutrient requirement
has been compromised in feed
formulation.
We need to have a clear
understanding on the species
specific trace minerals requirements in aquaculture.
As the aquafeed formulation
has been shifting to dietary plant
protein from fishmeal, emphasis
should also be placed to revise
the inclusion level of as well as
efficacy of new perspective to
ensure the bioavailability of trace
mineral.
At this point, we must appreciate what is known today and try
further improving our knowledge
on the significance of dietary trace
minerals in fish nutrition and health.

Acknowledgement:
I wish to thank my PhD student,
Kumar Katya for his input in
articulating this article.
May-June 2014 | INTERNATIONAL AQUAFEED | 23

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This digital re-print is part of the May | June 2014 edition of International
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online magazine on our website, and as an archive of individual features on
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I N C O R P O R AT I N G
F I S H FA R M I N G T E C H N O L O G Y

Assessing the potential

of polychaete meal in
shrimp feeds

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Contact the International Aquafeed Team

Subscribe to International Aquafeed

Prospects on dietary trace minerals

aquafeeds & aquaculture

Use of a heat-stable
protease in salmonid feeds
experiences from Canada and Chile

Fish Farming Technology supplement

Nets and cages

Recirculating aquaculture
systems

Vo l u m e 1 7 I s s u e 3 2 0 1 4 -

m AY | J u N e

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