Int.J.Curr.Microbiol.App.

Sci (2018) 7(12): 2053-2074

International Journal of Current Microbiology and Applied Sciences

ISSN: 2319-7706 Volume 7 Number 12 (2018)
Journal homepage: http://www.ijcmas.com

Review Article https://doi.org/10.20546/ijcmas.2018.712.236

An Evidential Review on Potential Benefits of Enzymes in

Aqua Feed Industry

N. Felix, E. Prabu*, B. Kannan and K. Manikandan

Directorate of Incubation and Vocational Training in Aquaculture, ECR-Muttukadu, Chennai,

Tamil Nadu Dr. J. Jayalalithaa Fisheries University, India

*Corresponding author

ABSTRACT

Enzymes are basically a type of protein in biological systems. They are generally used as
catalysts in order to catalyse the rate of reaction. Feeding these enzymes in the aquaculture
Keywords sector has some nutritional advances. Application of enzymes reducing the effects of anti-
nutritional factors, improves the dietary energy resulting in better performance of
Enzyme, Fish meal, fish/shrimps. Feed enzymes in the form of granules help enzymes to stay for longer time
Phytase, Protease, durations and are suitable for pelletisation process. Efficiency of feeds needs to be at
Xylanase, Plant maximum for economical operations. There are various kinds of enzymes which include
based ingredients
phytase, xylanase, cellulase, lipase, protease, amylase and many more which can increase
Article Info the nutrient availability, nutrient absorption during digestion, increase the rate of fish
growth and assist survival of fish in early stages of life. In addition, it makes the feeds
Accepted: more economical. Enzyme application may give a solution of high larval mortality of
15 November 2018 aquatic animals. Feeding larvae with enzymes would be beneficial. Enzymes play a
Available Online:
significant role in formulating cost effective, high quality and eco-friendly aqua feeds. At
10 December 2018
present, the use of enzymes in aqua feeds can reduce use of fishmeal which ultimately
reduces the cost of fish production. This may help to reduce the demand for fishmeal from
the aquaculture sector in coming years.

Introduction more meat and dairy foods. However, the

FAO data showed that world per capita meat
The United Nations Food and Agriculture consumption is increasing only for chicken
Organization (FAO) projected that world and fish. As the conversion of feed to edible
population will increase from the current 7.5 meat from fish is the most efficient for all
billion to 9.1 billion by 2050 (FAO, 2009). A animals farmed for meat, aquaculture is
significant increase in food production will be potentially the most viable source of future
required to feed this population growth and protein to meet global needs.
the FAO in its report on ―How to feed the
world in 2050‖ estimated that food production Economical fish and shrimp production
in developing countries will need to double requires maximum nutritional efficiency from
(FAO, 2009). Human diets are also shifting to feed. The main issue in aquaculture revolves

2053
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

around fish meal reduction in feeds and fish al., 2014). The major high protein plant
oil substitution in high energy diets. By 2030, ingredients in Norwegian salmon diets are soy
aquaculture production will contribute 62% or protein concentrate (24 %) and wheat/wheat
93.6 million tonnes to global seafood gluten (17 %) (Ytrestoyl et al., 2014), but
production (The World Bank, 2013). The increased use of other plant ingredients have
pattern of FM use has shifted nearly to be considered for further growth in the
exclusively to aqua feed production from aquaculture production.
livestock (Hardy 2010). Aquaculture
consumed 3.72 million tonnes or 60.8% of Challenges with plant-based ingredients
total FM produced (Tacon et al., 2011) and
0.78 million tonnes (73.8%) of global fish oil The most important challenges with plant
(FO) in 2008 (FAO 2012), at the expense of products as protein sources in feeds for fishes
the livestock sectors which have continued to particularly for carnivorous fish are: low level
reduce their usage of these marine of protein, low digestibility, high level of
commodities. By 2012, aquaculture’s fish carbohydrates, adverse amino acid profile,
meal consumption rose to 68% while FO other nutrients and the presence of anti
usage remained the same (74%) (Tacon and nutritional factors (Gatlin et al., 2007;
Metian, 2015). Despite efforts to improve fish Sorensen et al., 2010). Poor amino acid
meal availability and quality, global fish meal composition and unfair nutrient composition
production has remained static (5 – 7 million can be balanced by combining ingredients of
tonnes) year over year due to fully/over- different origin and use of additives such as
exploited fisheries while the production of amino acids, vitamins and minerals. (Sorensen
cereal grains and oil seeds are trending et al., 2010).
upwards at 2.9 billion and 574.1 million
tonnes respectively (USDA 2015). Further Lower digestibility
growth in the aquaculture production can
therefore not depend on an increase in the Nutrient digestibility of plant-based
catch volume of wild fish, but must rely on a ingredients is a critical component in
further increase in the use of alternative feed determining the potential of raw feedstuffs for
resources. The main source of plant based inclusion in fish feed. Digestibility refers the
protein aquatic feed includes soybean meal, amount of the nutrients/energy in the ingested
corn gluten meal, sunflower meal, feed that is not excreted by the animal (NRC
canola/rapeseed meal, peas and lupins. 2011). It is essential for optimising inclusion
Soybean meal having highest proportion of levels and minimising resource waste.
plant protein in fish diets owing to high yield,
relatively high crude protein content and easy Compared to FM, plant-based ingredients
and round the year availability. Nutritionists have relatively lower digestibility. This is due
are investigating the ways of utilising proteins to structural components (cellulose,
of plants origin, since they are cheaper, readily hemicellulose etc.) and metabolites (ANFs)
available, and easily accessible than animal which interfere with the animal’s digestive
protein sources. Plant ingredients have so far metabolism, lowering dietary nutrients
been the most cost efficient alternative, and absorption. Consequently, the nutritive value
cite an example, feeds for Norwegian farmed of a feedstuff also includes its nutrient and
salmon have changed from a marine based energy bioavailability (Altan and Korkut,
diet (90 % marine ingredients) to a plant based 2011).
diet (30 % marine ingredients) (Ytrestoyl et

2054
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

Anti-nutritional Factors (ANFs) configuration with six phosphate ester bonds

(Haros et al., 2005; Kumar et al., 2012).
Plants commonly synthesize metabolites of
low and high molecular weight called Phytate can strongly chelate with cations such
antinutritional factors as a defence mechanism as calcium, magnesium, zinc, copper, iron and
against herbivores (Khokar and Apenten, potassium to form insoluble salts. This
2003). ANFs are classified as endogenous adversely affects the absorption and digestion
compounds found in all plant-based of these minerals in fish (Papatryphon et al.,
ingredients which may negatively influence 1999). Around 50% to 80% of the total
feed intake, nutrient digestibility and phosphorous content in plant seeds is stored in
utilisation, growth, affect the function of the form of phytate (Ravindran et al., 1995).
internal organs and alter disease resistance Phosphorus in this form is generally not
(Krogdahl et al., 2010). They include, but are bioavailable to monogastric animals (human,
not limited to, phytases, Protease Inhibitors dogs, pigs, birds) and also to agastric animals
(PIs), Non-Starch Polysaccharides (NSPs) because they lack the intestinal digestive
(cellulose and hemicellulose), saponins, enzyme, phytase, required to separate
tannins, haemagglutinins or lectins, gossypols phosphorous from the phytate molecule
and cyanogenic glycosides (Soetan and (Jackson et al..1996). As a consequence of low
Oyewole, 2009). The structure and chemical digestibility of phytate by fish, most of the
composition, specifically heat-sensitivity, of phytate-P ends up being excreted into the
ANFs can determine which physical or water and may cause algal bloom pollution
chemical processes may be effective in (Baruah et al., 2004). Moreover phytate can
reducing their biological effects in animals also integrate with cation groups on protein,
(Khokar and Apenten, 2003). ANFs can be amino acids, starch and lipids in feedstuff
removed or inactivated by selective breeding, reducing the digestibility of these nutrients in
genetic modification, heat treatment or fish, poultry and pig. Phytate depresses protein
extraction (extrusion, pelleting, alcohol and amino acid digestibility and utilisation
extraction), or through supplementation efficiency in fish and other higher animals.
(enzyme, mineral, etc.) (Krogdahl et al., 2010) The concentration of phytate and phytase in
(Table 1). the feedstuffs varies considerably. Phytate
constitute between 0.7% and 2% of most
Phytate binds naturally occurring plant P cereal grains and oilseeds (Adeola and Sands,
making it unavailable to monogastrics and 2003). In general plant derived fish feed
impairs mineral absorption; NSP (soluble and ingredients such as soybean meal, rapeseed
insoluble) interferes with digestive processes meal, and sesame meal contain 1.0-1.5%, 5.0–
limiting nutrient uptake while PIs depress the 7.5% and 2.4% phytate respectively (Francis
digestion of protein, hindering amino acid et al., 2001) (Fig. 1).
absorption (Krogdahl et al., 2010).
Non-starch polysaccharides
Phytate-phosphorus
Dietary fibre is the portion of plant nutrient
Phytate is the primary storage form of P in containing lignin and polysaccharides
many plants accounting for 0.4 – 6.4% by (cellulose and hemicellulose) (McDonald et
weight and 60 – 90% of total P (Khokar and al., 2002; NRC, 2011). NSPs are,
Apenten, 2003). Phytate consists of an inositol hemicellulose, a complex group of
group, hexahydrocyclohexane in a chair polysaccharides (with the exception of starch)

2055
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

containing several hundred linked monomers that catalyse the hydrolytic cleavage of
of hexoses and pentoses (Sinha et al., 2011). specific peptide bonds in their target proteins
The main constituents are rhamnose, (Habib and Fazili, 2007). PIs are therefore
arabinose, xylose, glucose, galactose, proteins that form complexes with specific
mannose, glucuronic acid and galacturonic proteases (e.g. trypsin, chymotrypsin, etc) and
acid. Arabinoxylans (the arabinose and xylose suppress their activity along the GI trace
fractions) make up 60 – 70% of the endosperm (Krogdahl et al., 2010). In essence Protease
wall in most cereals with the exception of rice Inhibitors are natural anti-metabolic proteins
and barley where the percentages are 40% and which interfere with the digestive processes
20% respectively. and protein utilization, similar to the effects
seen with phytate (Alarcon et al., 1999).
Soybean meal the most highly utilised plant-
based ingredient, contains significant amounts Protease Inhibitors are found in nearly all
of NSPs (Ogunkoya et al., 2006). Raw soya plants accounting for 1-10% of total protein
beans contain approximately 200 g kg-1 NSP and are abundant in storage organs such as
(Refstie and Svihus, 1999) and cereals 100 – seeds and tubers (Wait et al., 2009). PIs
200 g kg-1 of NSPs in soluble and insoluble represent 6% of the protein present in soybean
forms (Castanon et al., 1997). RB contains and despite the efficiency of processing,
approximately 20 – 25% NSP which consist of residual levels may remain (Mikie et al.,
equal portions of cellulose and arabinoxylans 2009). Although some PIs are heat-labile and
(Choct, 1997). Arabinoxylans are also the can be eliminated using thermal treatments
major NSP in maize (NRC, 2011) (Fig. 2). (i.e. pelleting), some researches argue that
technological treatments do not always
Unlike the structure of starch, NSPs are guarantee elimination of trypsin inhibitor (a
composed of different monomers linked by β- type of serine protease inhibitor) in feeds
glycosidic bonds. The digestion of starch is (Lopez et al., 1999). However, other studies
facilitated by α-amylase, α-glucosidase and have confirmed that heat treatment typically
oligo-1,6-glucosidase, specialized enzymes for used in the extrusion process (>120o C) for
hydrolysing α-glycosidic bonds (Sinha et al., fish feed may be sufficient to inactivate most
2011). of the trypsin inhibitor activity in untreated
SBM (Romarheim et al., 2005)
In herbivores and some omnivores, the
activities of these enzymes range from high to Enzymes
medium, negating the need for exogenous
additives. Monogastrics, however, do not Enzymes are basically a type of protein in
produce enzymes such as –xylanase or – biological systems. They are generally used as
glucanase that can hydrolyse the bonds found catalysts in order to catalyze the rate of
in NSPs (Sinha et al., 2011). reaction. Enzymes catalyze the reaction to
convert complex substances into absorbable
Protease Inhibitors (PIs) substances. The catalysis reaction is very
specific to substrates. Feeding these enzymes
One of the main limitations of using high in the aquaculture sector has some nutritional
inclusions of plant-based feedstuff is their advances. Since last few years and will also
comparatively low quality protein content aid in reducing the effects of anti-nutritional
(López et al., 1999). The presence of PIs factors, improve the dietary energy resulting
reduce the activities of proteolytic digestive in better performance of fish/shrimps.
enzymes (i.e. protease). Proteases are enzymes
2056
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

Sources of enzymes Improves digestibility of nutrients

Enzymes are produced in all the living Reduces production of ammonia

organisms right from simple unicellular
organisms to complex higher forms of life.
Various microorganisms are involved in Phytases
enzyme production including bacteria from
Bacillus group, fungus from Aspergillus Phosphatases are a diverse group of enzymes
groups and yeasts. There are few microbes in that catalyse the hydrolysis of
the digestive tract of animals which are potent phosphomonoester bonds of various phosphate
in production of proteolytic enzymes and esters. Phytases are a sub-group of
cellulose. Incorporation of live microbes in phosphatases with specificity for hydrolysing
feed can produce enzymes. Microbial phytate into phosphoric acid and myo-inositol
fermentation technique is widely used in large phosphate (Haros et al., 2005), with complete
scale commercial applications. hydrolysis yielding one molecule of inositol
and six molecules of inorganic phosphate
Feed enzymes (Makhode, 2008). This action reduces the
chelation capacity of phytate (Kumar et al.,
Stability of enzymes is important in order to 2012).
incorporate them in feed. Heat stability is an
important parameter to be considered. Feed Phytase activity was first detected many
enzymes in the form of granules help enzymes decades ago in rice bran (Suzuki et al., 1997).
to stay for longer time durations and are Warden and Schaible (1962) are the earlier to
suitable for pelletization process. Efficiency of verify that exogenous phytase improve
feeds needs to be at maximum for economical phytate-P use and bone mineralization in
operations. There are various kinds of poultry. However, before 1990s, the
enzymes which include phytase, xylanase, application of phytase has mainly been
cellulase, lipase, protease, amylase and many confined to poultry and swine to improve
more which can increase the nutrient utilization of plant P. Initial commercial
availability, nutrient absorption during phytase, Natuphos was created from
digestion, increase the rate of fish growth and Aspergillus niger and was released in market
assist survival of fish in early stages of life. In in 1991 (Selle and Ravindran, 2007).
addition, it makes the feeds more economical.
Enzyme application may give a solution of Following the prologue of commercial
high larval mortality of aquatic animals. phytase, more emphasis were given to
Feeding larvae with enzymes would be evaluating the effects of supplemental phytase
beneficial. on nutrient utilization and growth of common
aquaculture species such as rainbow trout.
Advantages of feed enzymes (Forster et al., 1999), common carp (Cyprinus
carpio L.) (Schaffer et al., 1995), channel
catfish (Ictalurus punctatus) Li and Robinson,
Aid in improvement of digestion and 1997), Africian catfish (Clarias gariepinus)
absorption of nutrients such as fat and proteins (Van Weerd et al., 1999). Atlantic salmon
Improves metabolizable energy of diet (Salmo salar) (Storebakken et al., 2000),
stripped bass (Morone saxatilis) (Papatryphon
Lead to increased feed intake, gain in weight et al., 1999), and Nile tilapia (Oreochromis
niloticus) (Liebert and Portz, 2005).

2057
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

Phytase application in aquaculture concentration of bone Ca and P compared

with the non-supplemented group. These
Enhancement in phosphorus bioavailability results were similar to those observed for rohu
(Baruah et al., 2005), common carp (Schafer
Various scientists around the world reported a et al., 1995), and other fish species
positive effect of phytase supplementation on (Storebakken et al., 1998; Papatryphon et al.,
total P availability in fish. The following table 1999; Yan and Reigh, 2002; Debnath et al.,
3 shows that the bioavailability of P when 2005b; Liebert and Portz, 2005). From these
phytase is added in the feed ingredients for studies it can be concluded that bone ash and
different fishes. bone P are sensitive indicators of the P status
in fish. This is because the P requirement for
It was seen that exogenous phytase was maximum bone mineralization is greater than
substantially efficient in enhancing the maximum body weight gain. Insufficient P
bioavailability of P and thus reducing the intake leads to the mobilization of P from the
amount of faecal-P. Supplementation of bone and transfer to soft tissues and metabolic
phytase in fish feed reduces the phosphate processes (Baeverfjord et al., 1998). Phytase
load in water from fish and ultimately supplementation results increment in bone ash
prevents phosphate induced algal bloom in fish feed that is an indication of the
contamination. Any reduction in P excreted by increased mineral bioavailability in fishes
fish and other animals is of benefit to both the (Baruah et al., 2005; Debnath et al., 2005).
environment and sustainable production.
Phytase supplementation also enhances
Enhancement of bioavailability of other digestibility of minerals which are bound to
nutrients and minerals phytate. Addition of phytase in a semi-purified
diet containing 50% soybean meal in rainbow
The concentration of minerals in plasma, bone trout significantly improved the apparent
and whole body will be increased by the digestibility of Zn (Cheng et al., 2004).
addition of phytase in fish feeds (Jackson et Moreover, dietary phytase have been shown to
al., 1996; Van Weerd et al., 1999; increase the apparent availability of protein,
Papatryphon and Soares, 2001; Debnath et al., ash, Ca, Cu, Mg, Fe, Sr and Zn in low ash
2005; Liebert and Portz, 2005). diets while little effect in high ash diets
Supplementation of phytase at a level of 1000 (Sugiura et al., 2001).
FTU/kg diet was sufficient to enhance Ca, Mg
and Mn content of bone in channel catfish, Cheng and Hardy (2004) reported that graded
and addition of phytase at a level of 8000 level of phytase inclusion in the rainbow trout
FTU/kg feed significantly increased the diet did not affect body composition; whereas,
bioavailability of naturally occurring Zn from it was effective in releasing most minerals and
feed (Yan and Reigh, 2002). Phytase trace mineral. This result showed that
supplementation in rainbow trout increased supplementation of trace minerals in rainbow
the apparent absorption of Ca, Mg, Cu, Fe, Sr trout diets can be reduced when phytase is
and Zn in low-ash soybean meal diet (Sugiura added in the diet. Schafer et al., (1995)
et al., 2001). Baruah et al., (2005) conducted observed that P excretion was lower by 30%
an experiment on rohu fingerlings and found on feeding a diet supplemented with phytase
that Phytase-supplemented groups in general compared to a diet supplemented with mono
recorded significantly (p < 0.05) higher calcium phosphate.
percentage of bone ash and also higher

2058
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

Enhancement of protein and amino acid amino acid bioavailability. Research

digestibility conducted on rainbow trout by Predergast et
al., (1994) and Teskeredzic et al., (1995)
Nonselectively phytase binds with proteins showed that pre-treatment of rapeseed protein
and inhibits the activities of pepsin, trypsin concentrate with the enzyme phytase did not
and alpha-amylase (Liener, 1994) as well as to improve the protein utilisation by rainbow
decrease protein digestibility. De- trout. Similarly, no positive effect of phytase
phytinization of dietary phytate by exogenous on protein digestibility could be noted in
phytase accounts for increased protein rainbow trout (Lanari et al., 1998), Atlantic
utilisation in common carp (Schafer et al., salmon (Storebakken et al., 1998) and striped
1995), Atlantic salmon (Storebakken et al., bass (Papatryphon et al., 1999)
1998; Sugiura et al., 1998), Seabass (Oliva-
Teles et al., 1998), Tilapia (Heindl, 2002) and Similarly Riche et al., (2001) reported that
pangus (Debnath et al., 2005b) by corrupting Nile tilapia offered diet with and without
the pre-formed phytate–protein complex. phytase showed no difference in protein
Forster et al., (1999) assessed the potential of utilisation, and also concluded that the
using dietary phytase to improve the nutritive available methionine and lysine decreased
value of canola protein concentrate diets for with increasing incorporation of phytase pre-
rainbow trout. Similarly, chemical and treated soybean meal. Phytase addition in
enzymatic processing of canola meal poultry, pigs and swine diets also showed
efficiently lowered most of the anti-nutritional conflicting results as observed for fish. The
factors in rainbow trout. The digestibility and probable reason for the neutral and/or negative
nutritional value of expeller and solvent- interaction of phytase and amino acids is that
extracted Australian canola meals when removal of phytate may increase the efficiency
included in the diets of juvenile red seabream of other anti-nutritional factors and protect
(Pagrus auratus) was comparable to those of amino acids from degradation, or decrease
the fishmeal (Glencross et al., 2004). 6.6% leaching of water soluble components (Cao et
phytase supplementation of 500 FTU/kg diet al., 2007). More research is needed to obtain a
improves digestibility of crude protein in better insight into the mechanisms for the
Crucian carp (Lie et al., 1999). phytase-protein interaction and availability of
proteins and amino acids.
Phytase supplementation in expelled soybean
diet of rainbow trout increased ADC of amino Enhancement of growth performance
acid significantly compared to raw soybean
but had no significant effect when added in Supplementation of phytase-containing diets
extruded soyabean (Cheng and Hardy, 2003). neutralises the negative effects of phytate and
Spraying soybean meal-based diets with increases growth in fish. Positive impact of
phytase improves protein digestibilities in phytase on growth of fish has been reported by
rainbow trout (Vielma et al., 2004). Phytase a number of authors: Jackson et al., (1996) in
supplemented diet in pangus increased channel catfish, Vanweerd et al., (1999) in
apparent net protein utilisation (Debnath et al., African catfish, Papatryphon and Soares
2005) and apparent protein digestibility and (2001) in striped seabass, Vielma et al., (2000)
were significantly (p<0.01) higher at a in rainbow trout, Liebert and Portz (2005) in
minimum supplement of 500 FTU/kg or tilapia, Debnath et al., (2005) in pangus.
higher in contrast to diet without phytase. Nwanna et al., (2005), in common carp and
There is discrepancy among authors for the Baruah et al., (2007a) in rohu. These authors
positive impact of phytase on protein and have demonstrated phytase hydrolysis in
2059
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

plant-based diets by phytase and improvement conditions, (Baruah et al., 2007).

of fish growth and mineralization. Diet Supplementing exogenous microbial phytase
containing 250 FTU phytase per kg increases in feed ration exhort an enhancement in
the feed intake and increases the weight than growth rate and performance which could be
the control diet containing no phytase (Li and attributed to various factors, in individual and
Robinson, 1997). Increase in weight gain from combine form namely better bio-availability
243 to 459% in rainbow trout fed soybean of phosphorous (Rodehutscord and Pfeffer
meal-based diets with phytase and 1995; Vielma et al., 2000; Baruah et al., 2007)
phosphorous supplementation (Vielma et al., and minerals (Vielma et al., 2004; Debnath et
2004). Similar results were reports in al., 2005b), improved protein digestibility
salmonids (Sugiura et al., 2001). Nwanna and (Vielma et al., 2004; Debnath et al., 2005a;
Schwarz (2007), Nwanna et al., (2007) found Liebert and Portz 2005; Baruah et al., 2007a)
better growth observed in common carp fed a and increased absorption of nutrients owing to
diet (incubated plant feed ingredients) well functioning of the pyloric caeca region of
containing phytase than another diet (without the intestine (NRC, 1993).
incubated plant feed ingredients) with and
without phytase. This is probable because Reduction in pollution from aquaculture
incubation process reduce phytate content of operation
feed improve phosphorous and mineral usage
as compared to untreated diet. The optimal Discharge of high levels of soluble P from fish
growth of Nile Tilapia is achieved by phytase culture systems into open water environment
supplementation at 750-1250 FTU/kg in plant- stimulate phytoplankton growth, resulting in
based diets (Liebert and Portz, 2005). wide fluctuations in dissolved oxygen
Addition of phytase at 1500 FTU/kg diet in concentrations (Li et al., 2004). Many studies
contrast to no inclusion of phytase enhanced have reported a clear effect of phytase
the weight gain of rainbow trout (Vielma et supplementation in reducing P excretion from
al., 2001). No considerable effect of phytase fish. Total phosphorous effluent was
supplementation was noticed on performance significantly lowered when fish cultured with
of large sized rainbow trout fed diet a diet enriched with phytase (200 FTU/kg) (Ai
supplemented with phytase at 1000 FTU/kg et al., 2007). Similarly, soybean meal based
(Vielma et al., 2000) (Fig. 3). diets supplemented with phytase decreased the
excretion of phosphorous from red sea bream
No effect on growth performance, protein and maximum reduction was reported at 2000
digestibility, energy retention on phytase FTU/kg feed (Biswas et al., 2007b).
addition in the diet of sea bass (Olivia-Teles et Comparable results were observed in rainbow
al., 1998). Forster et al., (1999) and Sajjadi trout (Sugiura et al., 2001). Faecal waste of P
and Carter (2004) did not report any in rainbow trout was reduced by phytase
improvement in the growth of rainbow trout supplementation in soybean protein
and Atlantic salmon when fed with canola concentrate diet (Vielma et al., 1998) and a
protein concentrate incorporated with phytase. significant decrease was noticed when
Similarly Masumoto et al., (2001) and Yoo et practical feed supplemented with phytase at a
al., (2005) reported no effect of dietary level of 2000 FTU/kg was fed (Vielma et al.,
phytase on weight gain of Japanese flounder 2001). Phosphorus concentration in faecal
and Korean rockfish (Sebastrers schlegeli). matter was reduced when trout were fed a diet
The discrepancy in above findings may be with phytase supplemented at 500 and 1000
associated with differences in their diet FTU/kg compared to non-supplemented feed
composition and also with different rearing (Verlhac et al., 2007). Soybean based phytase
2060
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

supplemented diet considerably lower Numerous studies have however recently

excretion of phosphorous compared to the shown beneficial effect from hydrolysed
fishmeal diet fed to Atlantic salmon products from NSP, so called prebiotics, on
(Storebakken et al., 2000). Phosphorus content fish growth and health (reviewed by Ringø et
of faeces was also reduced in Atlantic salmon al., (2010)). These could either be included in
fed a phytase supplemented diet (Sajjadi and the feeds as prebiotics or indirectly given to
Carter, 2004). Microbial phytase the fish by adding exogenous enzymes in fish
supplementation in the diets of juvenile catfish diets that hydrolyse NSP (Stone, 2003; Sinha
reduced the excretion of faecal phosphorous et al., 2011). In recent times, many researchers
about 60% (Li and Robinson, 1997). Many focussing on NSP enzymes in fish feeds.
studies suggest potential environmental These have been studied and utilised in swine
benefits to the extent of 30% to 40% reduction and poultry industry for several time (Khattak
in P excretion (Omogbenigun et al., 2003) et al., 2006). NSP-enzymes include
glucanases, pentosanases, cellulosases and
NSP-enzymes xylanases. These enzymes hydrolyze NSP to
products available for bacteria as prebiotics or
A greater concern is the high content of for the fish as digestible nutrients (Sinha et al.,
indigestible carbohydrates such as non-starch 2011). Supplementations of these have also
polysaccharides (NSP) which dilute the shown to improve protein utilization and
dietary energy and protein concentration and growth in fish (Ai et al., 2007; Jiang et al.,
reduce feed digestibility, content of anti- 2014) (Table 4).
nutritional factors that affects fish health,
nutrient utilization and growth, and reduced Xylanase is a class of enzymes that degrades
digestibility/bioavailability of nutrients due to linear polysaccharides, and breaks drown
extensive processing (Stone, 2003; Sørensen hemicelluloses that are the major component
et al., 2010). Therefore, processing are used to of the cell wall from plant (Ganguly et al.,
increase the protein content and reduce the 2013). This enzyme have proven to be
level of NSP in plant ingredients used in feeds especially efficient in maize-soy-based diet to
for carnivorous fish like Atlantic salmon broilers where the enzyme disrupts the plant
(Salmo salar). Genetical selection and cell wall that allows water hydration and
optimization of growing conditions can also entering of endogenous enzyme to act for a
be used to optimize nutrient content of plants. better digestion of starch and proteins (Sinha
et al., 2011). Xylanases are naturally produced
Non-starch polysaccharides (NSPs) can be in numerous yeasts, fungi and bacteria
water soluble or insoluble. Soluble NSPs such (Goswami and Pathak, 2003).
as arabinoxylans swell and form viscous gels Ronozyme®WX (1000 U xylanase/g) from
when hydrated in the intestine, thus preventing DSM Nutritional Products (Switzerland) has
secreted enzymes from reaching digestible been used in several fish experiments. Ai et
substrates, and impeding digested nutrients al., (2007) showed that Japanese seabass
from migrating to the gut wall for absorption. (Lateolabrax japonicas) at 6 g fed a diet of
Insoluble NSPs such as cellulose and lignin plant protein as soybean meal (170 g/kg),
induce a ―cage‖ effect, and nutrients are rapeseed meal (100 g/kg) and peanut meal
trapped within the folds of the NSP molecules. (100 g/kg), improved growth and protein
Ronozyme®WX (xylanase) works to reduce utilization, by inclusion of 800 mg/kg diet of
the viscosity of NSP gels, and breaks down Ronozyme®WX. There are also available
insoluble NSPs as well as improving commercial enzyme complexes where
assimilation of digested peptides and fats. xylanase is present in combination with other
2061
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

enzymes like proteases and NSP enzymes. Switzerland) is at the moment the best
These were tested in several fish studies with solution for improving protein digestibility
variable results. Tilapia fed diets with soybean available to the feed industry. Experiments
meal (170 g/kg), rapeseed meal (170 g/kg) and using an in vitro poultry gut model show
cottonseed meal showed improved growth, significant improvements in ingredient
feed conversion and endogenous enzyme digestibility when ProAct is provided on top
activities with increased inclusion (0, 1 and of endogenous digestive enzymes, and results
1.5 g/kg) of a commercial enzyme complex are not expected to be different with fish. The
(Yingheng Biotechnology, China) with adoption of protease by the aquafeed industry
xylanase (1600 U/g), protease and β-glucanase is just beginning, so there is not much
(Lin et al., 2007). The ingredients was mixed information available on the benefits of
and cold pelleted through an experimental protease.
feed mill. Shahsavai (2011) showed that
common carp (30 – 50 g) fed diets with wheat However, Dalsgaard et al., (2012) were able to
bran (340 g/kg), soybean meal (150 g/kg) and show a significant improvement in apparent
cottonseed meal (140 g/kg) supplemented with digestibility of soy (34% inclusion level in the
1, 2 and 3 g/kg diet of an enzyme complex feed) and a significant decrease in solid N
(Endofeed W, GNC Bioferm, Canada), with waste excretion when protease alone or
xylanase (≥1200 IU/g), β-glucanase, cellulase protease combined with xylanase was added to
and hemicellulase had no effect on feed rainbow trout feed. Plant ingredients such as
conversion and growth. Farmazyme® soy, rapeseed and canola contain trypsin
(Famavet, Turkey) a multi enzyme complex inhibitors that stop trypsin from cutting
containing fungal xylanase, glucanase and protein into peptides before further digestion
other enzymes have shown to improve growth by other proteases in the intestine.
and protein content in 46 g Africian Catfish
(Claris gariepinus) (Yildirin and Turan 2010). ProAct has been shown to digest trypsin
The enzyme complex was mixed with water inhibitor proteins, thus improving digestive
and a pulverized trout diet at 0, 0.25, 0.5 and function; It is less specific in selecting active
0.75 g/kg diet, and grinded with a 2 mm die sites on proteins for digestion than trypsin,
plate. Growth and protein content was hence it actually accelerates the initial stages
significantly improved at level of the enzyme of protein breakdown. In an experiment with
complex above 0.5 mg/kg diet. As mention tilapia with three different protein levels and
earlier, however, some of these herbivorous three different dosages of ProAct enzyme
freshwater species have naturally occurring (Verlhac and Diaz, 2012), apparent protein
enzyme producing yeasts in their gut, which digestibility was improved from 2–4% in a
improve the carbohydrate digestibility. 31% crude protein (CP) diet, and from 3-8%
Therefore, supplementation of enzymes may for 28 and 26% CP diets (Table 2), suggesting
perhaps have larger effects on carnivorous fish that in feeds with lower quality protein the
species. benefit of using protease may be greater
Protease, then, has a lot of potential to
Proteases improve digestibility of all types of protein
ingredients, and will assist nutritionists in
Digestibility of protein and amino acids in formulating feeds that are more digestible and
alternative ingredients of plant and animal less polluting, while at the same time offering
origin can be improved by adding protease the possibility of choosing less expensive
enzyme to feeds. ProAct protease (DSM, ingredients to control formulation costs.

2062
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

Table.1 Processing steps for removal/inactivation of ANFs (Nwanna 2007)

Anti-Nutrient Heat Sensitivity Extraction Other Treatment

Phytic Acid No No Phytase
Arabinoxylans No No Xylanase
(NSP)
Protease Yes No Protease
inhibitors
Hemagglutinin Yes No No
Saponin No Yes No
Phytoestrogen No Yes No

Table.2 List of anti-nutrients in plant sources

Antinutrients Chemical name Plant source Source

Phytic acid or Myoinositol Cereal and legumes (Khokar and
Phytate-P 1,2,3,4,5,6- Apenten, 2003)
hexakisdihydrogen
phosphate
Non-Starch e.g. Arabinoxylans Cereals (wheat, rye, (Sinha et al., 2011)
Polysaccharides (arabinose and barley, rice,
xylose) sorghum)
Protease e.g. Trypsin Most plants (Francis et al.,
Inhibitors inhibitor particularly 2001; Krogdahl et
legumes and al., 2010)
cereals

Table.3 Total P and phytate-P in common plant-based ingredients. Source: Kumar et al., (2012)
and Ravindran et al., (1994)

Ingredients Total P Phytate-P Proportion of Phytate-P

(g kg-1) (g kg-1) in Total P (%)
Maize 2.40 2.05 85.4
Corn 2.50 1.70 73.0
Rice 1.20 0.80 65.0
RB 17.51 15.83 90.2
Soya bean 5.55 3.08 55.5
SBM 6.66 4.53 68.3
Cassava 1.60 0.40 25.0

2063
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

Table.4 NSP comparison of major plant-based ingredients (in g kg-1), NRC (2011)

Ingredients Total NSP Arabinoxylansa Other fractions b

Rice Bran 218 85 133
Corn 81 52 29
Maize 97 52 45
Soya beans 192 47 145
SBM 196 42 154

Table.5 Commercially available microbial phytases (Sources: Hou, 2001: Srefan et al., 2005;
Cao et al., 2007)

Company Country Phytase Source Production Strain Trademark

AB Enzymes Germany Aspergillus awamari Trichoderma reesei Finase

Aiko Biotechnology Finland A.oryzoe A.oryzoe SP, TP, SF

Alltech USA A.niger A.niger Allzyme

Phytase

BASF Germany A.niger A.niger Natuphos

Biozyme USA A.oryzoe A.oryzoe AMAFERM

DSM USA P.Lyci A.oryzoe Bio-feed

phytase

Fermic Mexico A.oryzoe A.oryzoe Phyzme

Finnfeeds Finland A.awamari T.reesei Avizyme

international

Genencor USA Penicillium Penicillium Rovabio

International simplicissimum funiculosum

Roal Finland A. Awamari T.reesei Finase

Novozymes Denmark A.oryzoe A.oryzoe Ronozyme

2064
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

Table.6

References Fish Feed ingredients P availability Phytase dose P availability (%)

(plant protein sources (%) without FTU Kg diet with phytase
phytase

Rainbow trout (Oncorhynchus mykiss)

Riche and Canola meal, solvent extracted 4.8;(-13.4)8.4; 3.8 x 106 46.2; 46.6; 64.4;
Brown (1996) soybean meal, full fat soybean, 22.1; 30.7; NA 75.6; 76.8; 56.3
peanut meal, corn gluten meal,
cotton seed meal, canola meal,
Barley, wheat
Cherg and Canola meal, Barley, Wheat 12.2; 79.4; 61.6 500 41.8; 82.7; 64.6
Hardy (2002)
Cherg and Raw soybean, expelled 21.2; NA; 12.5 750; 200; (200, NA; 31.7; (81.3. 92.2,
Hardy (2003) soybean; Extruded full fat 400, 600, 800, 89.7, 95.2, 93.9,)
soybean 1000)
Vielma et al., Rapeseed meal, soybean meal; (-1.0); 48.3; 61; 750 53.8; 85.2, 118; 45.7,
(2006) corn gluten meal, sunflower (-0.9); 45.0; 72; 846
meal; 65.2
Verlac et al., Soy protein concentrate; Pea 29.9; 74.1; 47.8 750 46.9; 80.3; 69.9
(2007) meal, Faba bean meal
Nile Tillapia (Oreochromis niloticus)
Verilac et al., Soybean meal. Palm kernel 47.9, 25.5; 35.,; 750 76.9; 50.4; 59.5; 58.3;
(2007) cake, rice bran, corn, cassava 23.6; 72.4 92.6
Sea bass (Dicentrarchus labrax)
Papatryphon Isolated soy protein; soybean 48; 59; 52; (-10) 1000 74; 87; 70; 11
and Soares meal, corn gluten mean, wheat
(2001) middings

Fig.1 Chemical structure of phytate-phosphorus showing its chair-like conformation. Source:

Adeola and Sands, (2003)

2065
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

Fig.2 Chemical structure of arabinoxylan [Source: Sinha et al., (2011)]

Fig.3 Hydrolysis of phytate by phytase. Source: Kumar et al., (2012)

2066
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

Benefits of combining enzymes higher profit margin (Chesson, 1993). In other

words, they must somehow improve upon
The cooperativity of enzymes to degrade least-cost formulation by lowering input cost
feedstuff and their interactions require much while maximizing outputs in terms of animal
research. The benefits of combining phytases performance, health and cost to produce one
and xylanases have been demonstrated to unit of animal protein. Economic benefits of
some extent in broilers (Bedford, 2000). using phytase are by far more straight forward
Several enzyme companies (Novozyme/Royal than those of xylanases and proteases. Phytase
DSM, Altech, Ameco-Bio & Cp., Canada delivers direct cost benefit by replacing the
Bio-Systems Inc etc.) are now producing need for inorganic phosphate (Bedford, 2000).
enzyme cocktails to improve, even further, The benefits of reducing P load and feed
the efficiencies of feed utilisation, particularly formulation cost are clear, and as a result
those with high inclusions of plant-based phytase is now considered a standard feed
ingredients, and the synergistic benefits for additive. Though most enzyme studies
animal performances. Combining enzymes acknowledge supplementation-related
may provide additional benefits, in that, formula cost savings, rarely are these figures
different enzymes act in different location published for reference (Table 6).
along the GI tract and target different
substrates (Walk, 2009). Enzyme research for the future

Considerable effects of multi-enzyme With the increasing use of more plant

supplementation on ADC of DM, CP, ingredients such as rice bran, wheat bran,
nitrogen free extract (NFE), P and GE in copra meal, and palm oil milling byproducts
SBM-based diets fed to rainbow trout in aqua feeds, there is merit in improving
(Ogunkoya et al., 2006). Using a similar digestion of plant cell walls to unlock
commercial enzyme complex, higher FI was valuable nutrients trapped inside cells. Cell
recorded with tilapia fed diets containing 0.15 walls of cereals (wheat, corn, barley, rice) are
g kg-1 but no difference were observed in mainly made of arabinoxylans and β-glucans,
protein, lipid and GE ADCs between whereas oilseed crops (soy, canola, rapeseed,
treatments (0, 0.15 and 1.0 g kg-1) (Lin et al., sunflower) are mainly xyloglucans and
2007). Khalafalla et al., (2010) also showed pectins. Feed enzymes that digest cellulose,
the addition of Amecozyme in diets at 0.5% xylans, glucans, mannans and pectins are now
and 1.0% enhanced the growth performance widely used in livestock and poultry feeds,
of O. niloticus fingerlings. Similarly, a but have yet to be applied to aqua feeds.
cocktail containing protease, xylanase, Adding phytase and xylanase together with
glucanse, lipase, amylase and cellulase was protease improves protein utilization the
used to supplement five grain diets fed to most, with phytase reducing phytic acid-
tilapia which improved fish performance, protein interactions, and xylanase improving
nutrient digestibility, carcass characteristics protein, peptide and amino acid migration in
and faecal recovery (Soltan, 2009) (Table 5). the intestine in feeds containing large
quantities of NSPs. Research is focussed to
Economic benefits of supplementation investigate combination of enzymes to
further improve the feed efficiency of fishes.
The use of enzymes must sufficiently
demonstrate substantial improvements in feed In conclusion, flourishing and sustainable
conversion or product quality to cover any aquaculture depends on efficiently viable and
adjustments in formula cost resulting in environmentally responsible aqua feeds. Feed
2067
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

is the major working cost involved in pp.1-12.

intensive farming of aquatic organisms. The Baruah, K., Pal, A.K., Sahu, N.P., Jain, K.K.,
major feed ingredient, fishmeal, is expensive Mukherjee, S.C. and Debnath, D., 2005.
and there is increasing competition with other Dietary protein level, microbial phytase,
livestock industries for the available supply. citric acid and their interactions on bone
Hence, researchers are focussing to find mineralization of Labeo rohita
alternatives to fishmeal. Substitution of (Hamilton) juveniles. Aquaculture
fishmeal with plant proteins supplemented Research, 36(8), pp.803-812.
with feed enzymes is an effective alternative Baruah, K., Sahu, N.P., Pal, A.K. and
in aqua feeds. Enzymes play a significant role Debnath, D., 2004. Dietary phytase: an
in formulating cost effective, high quality and ideal approach for a cost effective and
eco-friendly aqua feeds. At present, the use of low-polluting aquafeed. NAGA, World
enzymes in aqua feeds can reduce use of Fish Center Quarterly, 27(3-4), pp.15-
fishmeal which ultimately reduces the cost 19.
fish production. This may help to reduce the Baruah, K., Sahu, N.P., Pal, A.K., Debnath,
demand for fishmeal from the aquaculture D., Yengkokpam, S. and Mukherjee,
sector in coming years. S.C., 2007. Interactions of dietary
microbial phytase, citric acid and crude
References protein level on mineral utilization by
rohu, Labeo rohita (Hamilton),
Adeola, O. and Sands, J.S., 2003. Does juveniles. Journal of the World
supplemental dietary microbial phytase Aquaculture Society, 38(2), pp.238-249.
improve amino acid utilization? A Bedford, M.R., 2000. Exogenous enzymes in
perspective that it does not. Journal of monogastric nutrition—their current
Animal Science, 81, pp.78-85. value and future benefits. Animal Feed
Ai, Q., Mai, K., Zhang, W., Xu, W., Tan, B., Science and Technology, 86(1-2), pp.1-
Zhang, C. and Li, H., 2007. Effects of 13.
exogenous enzymes (phytase, non- Biswas, A.K., Kaku, H., Ji, S.C., Seoka, M.
starch polysaccharide enzyme) in diets and Takii, K., 2007. Use of soybean
on growth, feed utilization, nitrogen and meal and phytase for partial
phosphorus excretion of Japanese replacement of fish meal in the diet of
seabass, Lateolabrax japonicus. red sea bream, Pagrus major.
Comparative Biochemistry and Aquaculture, 267(1-4), pp.284-291.
Physiology Part A: Molecular & Cao, L., Wang, W., Yang, C., Yang, Y.,
Integrative Physiology, 147(2), pp.502- Diana, J., Yakupitiyage, A., Luo, Z. and
508. Li, D., 2007. Application of microbial
Altan, Ö. and Korkut, A.Y., 2011. Appearent phytase in fish feed. Enzyme and
digestibility of plant protein based diets Microbial Technology, 40(4), pp.497-
by european sea bass Dicentrarchus 507.
labrax L., 1758. Turkish Journal of CAO, L., WANG, W.M., YANG, C.T. and
Fisheries and Aquatic Sciences, 11(1). YANG, Y., 2007. Effects of Microbial
Baeverfjord, G., Asgard, T. and Shearer, Phytase on Phytate P Contents in Three
K.D., 1998. Development and detection Plant Ingredients [J]. Freshwater
of phosphorus deficiency in Atlantic Fisheries, 4, p.013.
salmon, Salmo salar L., parr and post- Castañón, M.L., Ordieres, A.M. and Blanco,
smolts. Aquaculture Nutrition, 4(1), P.T., 1997. Amperometric detection of

2068
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

ethanol with poly-(o- K.K., Yengkokpam, S. and Mukherjee,

phenylenediamine)-modified enzyme S.C., 2005. Effect of dietary microbial
electrodes. Biosensors and phytase supplementation on growth and
Bioelectronics, 12(6), pp.511-520. nutrient digestibility of Pangasius
Cheng, Z.J. and Hardy, R.W., 2004. Effects of pangasius (Hamilton) fingerlings.
microbial phytase supplementation in Aquaculture Research, 36(2), pp.180-
corn distiller's dried grain with solubles 187.
on nutrient digestibility and growth FAO 2009. The state of world fisheries and
performance of rainbow trout, aquaculture 2010. Food and
Oncorhynchus mykiss. Journal of Agricultural Organisation, Rome, p. 18-
Applied Aquaculture, 15(3-4), pp.83- 25.
100. FAO 2012. The state of world fisheries and
Cheng, Z.J., Hardy, R.W. and Usry, J.L., aquaculture 2010. Food and
2003. Effects of lysine supplementation Agricultural Organisation, Rome, p.
in plant protein-based diets on the Forster, I., Higgs, D.A., Dosanjh, B.S.,
performance of rainbow trout Rowshandeli, M. and Parr, J., 1999.
(Oncorhynchus mykiss) and apparent Potential for dietary phytase to improve
digestibility coefficients of nutrients. the nutritive value of canola protein
Aquaculture, 215(1-4), pp.255-265. concentrate and decrease phosphorus
Cheng, Z.J., Hardy, R.W., Verlhac, V. and output in rainbow trout (Oncorhynchus
Gabaudan, J., 2004. Effects of microbial mykiss) held in 11 C fresh water.
phytase supplementation and dosage on Aquaculture, 179(1-4), pp.109-125.
apparent digestibility coefficients of Francis, G., Makkar, H.P. and Becker, K.,
nutrients and dry matter in soybean 2001. Antinutritional factors present in
product‐ based diets for rainbow trout plant-derived alternate fish feed
Oncorhynchus mykiss. Journal of the ingredients and their effects in fish.
world aquaculture society, 35(1), pp.1- Aquaculture, 199(3-4), pp.197-227.
15. Ganguly, S., Dora, K.C., Sarkar, S. and
Chesson, A., 1993. Feed enzymes. Animal Chowdhury, S., 2013. Supplementation
feed science and technology, 45(1), of prebiotics in fish feed: a review.
pp.65-79. Reviews in Fish Biology and Fisheries,
Choct, M., 1997. Feed non-starch 23(2), pp.195-199.
polysaccharides: chemical structures Gatlin III, D.M., Barrows, F.T., Brown, P.,
and nutritional significance. Feed Dabrowski, K., Gaylord, T.G., Hardy,
milling international, 191(June issue), R.W., Herman, E., Hu, G., Krogdahl,
pp.13-26. Å., Nelson, R. and Overturf, K., 2007.
Dalsgaard, J., Verlhac, V., Hjermitslev, N.H., Expanding the utilization of sustainable
Ekmann, K.S., Fischer, M., Klausen, M. plant products in aquafeeds: a review.
and Pedersen, P.B., 2012. Effects of Aquaculture research, 38(6), pp.551-
exogenous enzymes on apparent 579.
nutrient digestibility in rainbow trout Glencross, B., Evans, D., Hawkins, W. and
(Oncorhynchus mykiss) fed diets with Jones, B., 2004. Evaluation of dietary
high inclusion of plant-based protein. inclusion of yellow lupin (Lupinus
Animal feed science and technology, luteus) kernel meal on the growth, feed
171(2-4), pp.181-191. utilisation and tissue histology of
Debnath, D., Pal, A.K., Sahu, N.P., Jain, rainbow trout (Oncorhynchus mykiss).

2069
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

Aquaculture, 235(1-4), pp.411-422. activity relations. Food Chemistry,

Goswami, G.K. and Pathak, R.R., 2017. 81(1), pp.133-140.
Microbial xylanases and their Krogdahl, Å., Penn, M., Thorsen, J., Refstie,
biomedical applications: a review. S. and Bakke, A.M., 2010. Important
International Journal of Basic & antinutrients in plant feedstuffs for
Clinical Pharmacology, 2(3), pp.237- aquaculture: an update on recent
246. findings regarding responses in
Habib, H. and Fazili, K.M., 2007. Plant salmonids. Aquaculture Research,
protease inhibitors: a defense strategy in 41(3), pp.333-344.
plants. Biotechnology and Molecular Kumar, V., Sinha, A.K., Makkar, H.P.S., De
Biology Reviews, 2(3), pp.68-85. Boeck, G. and Becker, K., 2012.
Hardy, R.W., 2010. Utilization of plant Phytate and phytase in fish nutrition.
proteins in fish diets: effects of global Journal of animal physiology and
demand and supplies of fishmeal. animal nutrition, 96(3), pp.335-364.
Aquaculture Research, 41(5), pp.770- Lanari, D., D'agaro, E. and Turri, C., 1998.
776. Use of nonlinear regression to evaluate
Haros, M., Bielecka, M., Sanz, Y., 2005. the effects of phytase enzyme treatment
Phytase activity as a novel metabolic of plant protein diets for rainbow trout
feature in Bifidobacterium. FEMS (Oncorhynchus mykiss). Aquaculture,
Microbiology Letters 247, 231–239. 161(1-4), pp.345-356.
Jackson, L.S., Li, M.H. and Robinson, E.H., Li, M.H. and Robinson, E.H., 1997. Microbial
1996. Use of Microbial Phytase in Phytase Can Replace Inorganic
Channel Catfish Ictalurus punctatus Phosphorus Supplements in Channel
Diets to Improve Utilization of Phytate Catfish Ictalurus punctatus Diets 1.
Phosphorus 1. Journal of the World Journal of the World Aquaculture
Aquaculture Society, 27(3), pp.309-313. Society, 28(4), pp.402-406.
Jackson, R.B. and Caldwell, M.M., 1996. Lie, G.X., Li, X.J. and Qiao, Z.G., 1999.
Integrating resource heterogeneity and Application of phytase in feed industry.
plant plasticity: modelling nitrate and Journal of Guangdong Feed, 12, pp.36-
phosphate uptake in a patchy soil 38.
environment. Journal of Ecology, Liebert, F. and Portz, L., 2005. Nutrient
pp.891-903. utilization of Nile tilapia Oreochromis
Khalafalla, M.M., Bassiouni, M., Eweedah, niloticus fed plant based low
N. and Elmezyme, H., 2010. phosphorus diets supplemented with
Performance of Nile tilapia graded levels of different sources of
(Oreochromis niloticus) fingerlings fed microbial phytase. Aquaculture, 248(1-
diets containing different levels of 4), pp.111-119.
Amecozyme. Journal of Agriculture Liener, I.E., 1994. Implications of
Research, 36(2), pp.111-122. antinutritional components in soybean
Khattak, F.M., Pasha, T.N., Hayat, Z. and foods. Critical Reviews in Food Science
Mahmud, A., 2006. Enzymes in poultry & Nutrition, 34(1), pp.31-67.
nutrition. Journal of Animal and Plant Lin, S., Mai, K. and Tan, B., 2007. Effects of
Sciences, 16(1-2), pp.1-7. exogenous enzyme supplementation in
Khokhar, S. and Apenten, R.K.O., 2003. Iron diets on growth and feed utilization in
binding characteristics of phenolic tilapia, Oreochromis niloticus x O.
compounds: some tentative structure– aureus. Aquaculture research, 38(15),

2070
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

pp.1645-1653. Oliva-Teles, A., 1998. Apparent digestibility

Makhode, M.L., 2008. Enzymatic and coefficients of feedstuffs in seabass
molecular characterization of phytase (Dicentrarchus labrax) juveniles.
producing yeasts isolated from soil in Aquatic Living Resources, 11(3),
the Limpopo Province (Doctoral pp.187-191.
dissertation). Olivia-Teles, A., PEREIRA, J. and Gouveia,
Masumoto, T., Tamura, B. and Shimeno, S., A., 1998. Utilization of diets
2001. Effects of phytase on supplemented with microbial phytase by
bioavailability of phosphorus in seabass. Aquatic Living Resources,
soybean meal‐ based diets for Japanese 11(4), pp.225-259.
flounder Paralichthys olivaceus. Omogbenigun, F.O., Nyachoti, C.M. and
Fisheries science, 67(6), pp.1075-1080. Slominski, B.A., 2003. The effect of
McDonald, P., 2002. Animal nutrition. supplementing microbial phytase and
Pearson education. organic acids to a corn-soybean based
NRC (National Research Council) 1993. diet fed to early-weaned pigs. Journal
Nutrient Requirement of Fish. National of Animal Science, 81(7), pp.1806-1813.
Academy Press, Washington, DC, USA, Papatryphon, E. and Soares, J.H., 2001. The
pp. 45-47 effect of phytase on apparent
Nwanna, L.C. and Schwarz, F.J., 2007. Effect digestibility of four practical plant
of supplemental phytase on growth, feedstuffs fed to striped bass, Morone
phosphorus digestibility and bone saxatilis. Aquaculture Nutrition, 7(3),
mineralization of common carp pp.161-168.
(Cyprinus carpio L). Aquaculture Papatryphon, E., Howell, R.A. and Soares Jr,
Research, 38(10), pp.1037-1044. J.H., 1999. Growth and mineral
Nwanna, L.C., Eisenreich, R. and Schwarz, absorption by striped bass Morone
F.J., 2007. Effect of wet-incubation of saxatilis fed a plant feedstuff based diet
dietary plant feedstuffs with phytases on supplemented with phytase. Journal of
growth and mineral digestibility by the World Aquaculture Society, 30(2),
common carp (Cyprinus carpio L). pp.161-173.
Aquaculture, 271(1-4), pp.461-468. Predergast, J., 1994. The forgotten agenda in
Nwanna, L.C., Fagbenro, O.A. and Adeyo, Somalia.
A.O., 2005. Effects of different Ravindran, V., Ravindran, G. and Sivalogan,
treatments of dietary soybean meal and S., 1994. Total and phytate phosphorus
phytase on the growth and mineral contents of various foods and feedstuffs
deposition in African catfish Clarias of plant origin. Food Chemistry, 50(2),
gariepinus. Journal of Animal and pp.133-136.
Veterinary Advances, 4(12), pp.980- Refstie, S., Korsøen, Ø.J., Storebakken, T.,
987. Baeverfjord, G., Lein, I. and Roem,
Ogunkoya, A.E., Page, G.I., Adewolu, M.A. A.J., 2000. Differing nutritional
and Bureau, D.P., 2006. Dietary responses to dietary soybean meal in
incorporation of soybean meal and rainbow trout (Oncorhynchus mykiss)
exogenous enzyme cocktail can affect and Atlantic salmon (Salmo salar).
physical characteristics of faecal Aquaculture, 190(1-2), pp.49-63.
material egested by rainbow trout Refstie, S., Svihus, B., Shearer, K.D. and
(Oncorhynchus mykiss). Aquaculture, Storebakken, T., 1999. Nutrient
254(1-4), pp.466-475. digestibility in Atlantic salmon and

2071
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

broiler chickens related to viscosity and Microbial phytase in poultry nutrition.

non-starch polysaccharide content in Animal Feed Science and Technology,
different soyabean products. Animal 135(1-2), pp.1-41.
Feed Science and Technology, 79(4), Sinha, A.K., Kumar, V., Makkar, H.P., De
pp.331-345. Boeck, G. and Becker, K., 2011. Non-
Riche, M., Trottier, N.L., Ku, P.K. and starch polysaccharides and their role in
Garling, D.L., 2001. Apparent fish nutrition–A review. Food
digestibility of crude protein and Chemistry, 127(4), pp.1409-1426.
apparent availability of individual Soetan, K.O. and Oyewole, O.E., 2009. The
amino acids in tilapia (Oreochromis need for adequate processing to reduce
niloticus) fed phytase pretreated the anti-nutritional factors in plants used
soybean meal diets. Fish Physiology as human foods and animal feeds: A
and Biochemistry, 25(3), p.181. review. African Journal of food science,
Rodehutscord, M. and Pfeffer, E., 1995. 3(9), pp.223-232.
Effects of supplemental microbial Soltan, M.A., 2009. Effect of dietary fish
phytase on phosphorus digestibility and meal replacement by poultry by-product
utilization in rainbow trout meal with different grain source and
(Oncorhynchus mykiss). Water Science enzyme supplementation on
and technology, 31(10), pp.143-147. performance, feces recovery, body
Romarheim, O.H., Aslaksen, M.A., composition and nutrient balance of
Storebakken, T., Krogdahl, Å. and Nile tilapia. Pakistan Journal of
Skrede, A., 2005. Effect of extrusion on Nutrition, 8(4), pp.395-407.
trypsin inhibitor activity and nutrient Sørensen, M., Penn, M., El-Mowafi, A.,
digestibility of diets based on fish meal, Storebakken, T., Chunfang, C.,
soybean meal and white flakes. Øverland, M. and Krogdahl, Å., 2011.
Archives of animal nutrition, 59(6), Effect of stachyose, raffinose and soya-
pp.365-375. saponins supplementation on nutrient
Sajjadi, M. and Carter, C.G., 2004. Dietary digestibility, digestive enzymes, gut
phytase supplementation and the morphology and growth performance in
utilisation of phosphorus by Atlantic Atlantic salmon (Salmo salar, L).
salmon (Salmo salar L.) fed a canola- Aquaculture, 314(1-4), pp.145-152.
meal-based diet. Aquaculture, 240(1-4), Storebakken, T., 2000. Soy products as fat
pp.417-431. and protein sources in fish feeds for
Sajjadi, M. and Carter, C.G., 2004. Effect of intensive aquaculture. Soy in animal
phytic acid and phytase on feed intake, nutrition, pp.127-170.
growth, digestibility and trypsin activity Storebakken, T., Kvien, I.S., Shearer, K.D.,
in Atlantic salmon (Salmo salar, L.). Grisdale-Helland, B., Helland, S.J. and
Aquaculture Nutrition, 10(2), pp.135- Berge, G.M., 1998. The apparent
142. digestibility of diets containing fish
Schäfer, A., Koppe, W.M., Meyer-Burgdorff, meal, soybean meal or bacterial meal
K.H. and Günther, K.D., 1995. Effects fed to Atlantic salmon (Salmo salar):
of a microbial phytase on the utilization evaluation of different faecal collection
of native phosphorus by carp in a diet methods. Aquaculture, 169(3-4),
based on soybean meal. Water Science pp.195-210.
and technology, 31(10), p.149. Storebakken, T., Shearer, K.D. and Roem,
Selle, P.H. and Ravindran, V., 2007. A.J., 1998. Availability of protein,

2072
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

phosphorus and other elements in fish National Plant Data Team, Greensboro.
meal, soy-protein concentrate and Van Weerd, J.H., K.H.A. Khalaf, F.J. Aartsen
phytase-treated soy-protein-concentrate- and P.A.T. Tijssen. 1999. Balance trials
based diets to Atlantic salmon, Salmo with African catfish Clarias gariepinus
salar. Aquaculture, 161(1-4), pp.365- fed phytase-treated soybean mealbased
379. diets. Aqua. Nutr. 5:135-142
Sugiura, S.H., Babbitt, J.K., Dong, F.M. and Van Weerd, J.H., Khalaf and T. Aartsen,
Hardy, R.W., 2000. Utilization of fish 1999. Balance trials with African catfish
and animal by‐ product meals in Clarias gariepinus fed phytase-treated
low‐ pollution feeds for rainbow trout soybean meal based diets. Aquacult.
Oncorhynchus mykiss (Walbaum). Nutr, 5, pp.135-142.
Aquaculture Research, 31(7), pp.585- Van Weerd, J.H., Khalaf and T. Aartsen,
593. 1999. Balance trials with African catfish
Sugiura, S.H., Gabaudan, J., Dong, F.M. and Clarias gariepinus fed phytase-treated
Hardy, R.W., 2001. Dietary microbial soybean meal based diets. Aquacult.
phytase supplementation and the Nutr, 5, pp.135-142.
utilization of phosphorus, trace minerals Verlhac, V., Vielma, J. and Gabaudan, J.,
and protein by rainbow trout 2007. The use of phytase in aquatic
[Oncorhynchus mykiss (Walbaum)] fed feeds. Aqua feed Horizons, VIctam
soybean meal‐ based diets. Aquaculture International 2007. Utrecht, the
Research, 32(7), pp.583-592. Netherlands (aquafeed.
Suzuki, Y.J., Forman, H.J. and Sevanian, A., com/docs/ah2007/VERLHAC. doc).
1997. Oxidants as stimulators of signal Vielma, J., Mäkinen, T., Ekholm, P. and
transduction. Free Radical Biology and Koskela, J., 2000. Influence of dietary
Medicine, 22(1-2), pp.269-285. soy and phytase levels on performance
Tacon, A.G. and Metian, M., 2015. Feed and body composition of large rainbow
matters: satisfying the feed demand of trout (Oncorhynchus mykiss) and algal
aquaculture. Reviews in Fisheries availability of phosphorus load.
Science & Aquaculture, 23(1), pp.1-10. Aquaculture, 183(3-4), pp.349-362.
Tacon, A.G., Hasan, M.R. and Metian, M., Vielma, J., Ruohonen, K., Gabaudan, J. and
2011. Demand and supply of feed Vogel, K., 2004. Top‐ spraying
ingredients for farmed fish and soybean meal‐ based diets with phytase
crustaceans: trends and prospects. FAO improves protein and mineral
Fisheries and Aquaculture Technical digestibilities but not lysine utilization
Paper, (564), p.I. in rainbow trout, Oncorhynchus mykiss
Teskeredžić, Z., Higgs, D.A., Dosanjh, B.S., (Walbaum). Aquaculture Research,
McBride, J.R., Hardy, R.W., Beames, 35(10), pp.955-964.
R.M., Jones, J.D., Simell, M., Vaara, T. Walk, C.L., Cowieson, A.J., Remus, J.C.,
and Bridges, R.B., 1995. Assessment of Novak, C.L. and McElroy, A.P., 2011.
undephytinized and dephytinized Effects of dietary enzymes on
rapeseed protein concentrate as sources performance and intestinal goblet cell
of dietary protein for juvenile rainbow number of broilers exposed to a live
trout (Oncorhynchus mykiss). coccidia oocyst vaccine. Poultry
Aquaculture, 131(3-4), pp.261-277. science, 90(1), pp.91-98.
USDA, N., 2015. The PLANTS Database Warden, W.K. and Schaible, P.J., 1962.
(http://plants. usda. gov, May 2011). Preliminary investigations concerning

2073
 Int.J.Curr.Microbiol.App.Sci (2018) 7(12): 2053-2074

utilization of phytin phosphorus by the Ictalurus punctatus fed an

chick. Poult. Sci, 41, p.1692. all‐ plant‐ protein diet. Journal of the
Xin, L., Hong-ying, H., Ke, G. and Ying-xue, World Aquaculture Society, 33(1),
S., 2010. Effects of different nitrogen pp.10-22.
and phosphorus concentrations on the Yildirim, Y.B. and Turan, F., 2010. Effects of
growth, nutrient uptake, and lipid exogenous enzyme supplementation in
accumulation of a freshwater microalga diets on growth and feed utilization in
Scenedesmus sp. Bioresource African catfish, Clarias gariepinus.
technology, 101(14), pp.5494-5500. Journal of Animal and Veterinary
Yan, W., Reigh, R.C. and Xu, Z., 2002. Advances, 9(2), pp.327-331.
Effects of fungal phytase on utilization Ytrestøyl, T., Aas, T.S. and Åsgård, T.E.,
of dietary protein and minerals, and 2014. Resource utilisation of
dephosphorylation of phytic acid in the Norwegian salmon farming in 2012.
alimentary tract of channel catfish

How to cite this article:

Felix, N., E. Prabu, B. Kannan and Manikandan, K. 2018. Need of Enzymes in Aqua Feeds.
Int.J.Curr.Microbiol.App.Sci. 7(12): 2053-2074. doi: https://doi.org/10.20546/ijcmas.2018.712.236

2074