Fish Physiol Biochem (2010) 36:1171–1180

DOI 10.1007/s10695-010-9395-6

Inhibitory effect of Pistia tannin on digestive enzymes

of Indian major carps: an in vitro study
Sudipta Mandal • Koushik Ghosh

Received: 26 June 2009 / Accepted: 22 March 2010 / Published online: 6 April 2010
Ó Springer Science+Business Media B.V. 2010

Abstract Aquatic weeds are one of the major on the elimination of tannin while incorporating feed
unconventional feed ingredients tested for aquafeed ingredients of plant origin in fish diets.
formulation. Tannin content in the water lettuce,
Pistia, has been quantified (26.67 mg g-1; dry Keywords Indian major carps
Tannin

weight) and graded levels of which (12.5–200 lg) Protease
Amylase
Lipase
Pistia
have been incorporated in the reaction mixtures to
evaluate any change in the in vitro activity of the
principal digestive enzymes from the three Indian
major carps (IMC), namely rohu (Labeo rohita), catla
(Catla catla) and mrigala (Cirrhinus mrigala). Result Introduction
of the experiment revealed that the Pistia tannin (PT)
significantly inhibit/lower the activities of the diges- The growing demand to substitute fish meal in
tive enzymes from three IMCs in a dose-dependant aquafeed has compelled to search for alternative less
manner, even at very low concentration. Significant expensive and protein-rich sources. Plant proteins are
variation in the reduction of the enzyme activities considered to be the most viable alternative in this
was noticed between the three fish species, as well as respect for economic fish production in most of the
between the three enzymes studied. Among the three developing countries (Wee and Wang 1987; Mukho-
species studied, digestive enzymes from L. rohita padhyay and Ray 1996; Becker and Makkar 1999).
were found to be the most sensitive to the PT, The use of plant-derived materials (e.g., aquatic
whereas enzymes from C. catla were found to be weeds, legume seeds, different types of oilseed cake,
comparatively least affected. On the other hand, leaf meals, leaf protein concentrates and root tuber
protease and lipase activities were comparatively meals) as fish feed ingredients is limited by the
more affected than the amylase activity. The results presence of a wide variety of antinutritional sub-
of the study suggest that more stress should be given stances (Francis et al. 2001). Even though possibilities
for uses of plant ingredients have been investigated
and achieved experimental success (Hasan et al. 1990;
Patra et al. 2002; Kalita et al. 2007). The water lettuce,
S. Mandal
K. Ghosh (&) Pistia, is an aquatic weed; infestation of which causes
Aquaculture Laboratory, Department of Zoology,
great problem in the aquaculture ponds. In compar-
The University of Burdwan, Golapbag, Burdwan 713104,
West Bengal, India ison with the terrestrial conventional vegetal meals,
e-mail: kghosh_bu@yahoo.co.in; kghoshbu@gmail.com the use of aquatic weeds like Pistia involves minimum

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1172 Fish Physiol Biochem (2010) 36:1171–1180

expenditure as it requires only the cost of labor for After soaking, the mixture was boiled for 30 min,
collection. Ray and Das (1995) have shown that Pistia cooled and centrifuged at 2,000 rpm for 20 min. The
has the potential to serve as one of such alternate supernatant was collected and used as tannin extract.
ingredient in fish feed. However, higher levels of The extracted tannin was not purified further.
incorporation of Pistia meal in aquafeeds were
associated with poor growth, feed conversion and Estimation of extracted tannin
protein utilization, which may be due to the interfer-
ence caused by the antinutrients therein. Tannins are Concentration of tannin in the extract was measured
one such naturally occurring plant polyphenols that by Folin–Denis method (Schanderi 1970) with minor
may interfere with the digestive processes (Liener modifications. The crude extract (0.2 ml) was diluted
1989). These are secondary compounds of various with 8.3 ml of distilled water and then mixed with
chemical structures which are generally divided into 0.5 ml of Folin–Denis reagent. The reaction mixture
hydrolysable and condensed tannins (Francis et al. was alkalinized by the addition of 1 ml of 15% (w/v)
2001). Experimental evidence has shown to induce sodium carbonate solution and kept in dark for
feed rejection by hydrolysable tannin when included 30 min at room temperature. The absorbance of the
in the feed of the common carp, Cyprinus carpio solution was read at 700 nm using spectrophotometer
(Becker and Makkar 1999). Poor growth performance (Shimadzu UV/VIS-1700), and the concentration of
has also been recorded in the same species as a result tannin in the extract was determined using pure
of feeding tannic acid incorporated diet (Hossain and tannic acid (MERCK, India) as standard.
Jauncey 1989). However, studies on the interaction of
tannin with fish intestinal enzymes are scanty. Maitra Experimental fish
and Ray (2003) demonstrated the inhibition of the
intestinal enzymes in Labeo rohita fingerlings to a Fingerlings of Labeo rohita, Catla catla and Cirrhi-
significant level by the tannin extracted from Acacia, nus mrigala were collected from three local compos-
though the effect of the hydrolysable tannin on the ite carp culture farms and acclimatized separately
digestive enzymes of the other carp species has not according to their source in glass aquaria (75 L) for
been worked out. Indian major carps might not share 10 days, during which fish were fed ad libitum with a
same pattern of inhibition with regard to the digestive diet containing approximately 40% crude protein
enzymes as they vary in feeding regime specificity having fish meal as the chief protein source. Average
and also in the enzyme activities (Jhingran 1997). weight of the fishes examined and their feeding habits
Therefore, the present study was undertaken to are presented in Table 1.
demonstrate the comparative effect of the hydrolysa-
ble tannin extracted from Pistia on the digestive Preparation of crude enzyme
enzymes of three Indian major carps, rohu, Labeo
rohita; catla, Catla catla and mrigal, Cirrhinus The experimental fish were weighed to the nearest
mrigala. gram on a single-pan top-loading balance, and the
anterior and middle intestinal parts were dissected out
on cooled plates and placed in prechilled Petri dishes.
Materials and methods Blood and other debris were washed out with chilled
phosphate buffer (0.1 mol L-1, pH 7.4) containing
Extraction of crude tannin 0.89% sodium chloride (phosphate buffered saline,
PBS). The cleaned intestinal parts were minced with
Tannin was extracted from Pistia leaves following scissors, and a 10% homogenate was prepared in the
the method described by Schanderi (1970). Pistia same buffer using tissue homogenizer (REMI, Model
leaves were collected from a local water body, oven RQ-127A2). The homogenate was centrifuged
dried (at 55 ± 5°C) for 48 h and finely powdered in a (REMI, Model No.C24) at 10,000 rpm for 30 min
mixer grinder (Remi Laboratory Blender). The pow- at 4°C. The supernatant was separated and used as
dered material (5 g) was mixed with distilled water enzyme extract. Preparation of enzyme extract and
(200 ml) and kept at room temperature for overnight. inhibition assay was done on the same day, and the

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Fish Physiol Biochem (2010) 36:1171–1180 1173

Table 1 Food habit and average weight of the fishes examined

Fish species Food habit Average weight (g) (±SD)

Catla catla Zooplanktophagous 17.67 ± 1.8

Labeo rohita Omnivorous, mostly plant matter 12.44 ± 1.67
Cirrhinus mrigala Detritivorous 10.11 ± 1.45
Values are mean ± standard deviation (SD) of nine specimens

extract was kept at 4°C in between. Protein content of 1 ml of ninhydrin reagent were added in a test tube
the extract was determined following the method and heated in a boiling water bath for 20 min, placing
described by Lowry et al. (1951) using BSA as a marble on top of each tube. After cooling to room
standard. Three specimens of each fish species from temperature, the developed color was read at 570 nm
each collection site were used for preparation of in a spectrophotometer (Shimadzu UV/VIS-1700).
enzyme extract for every replicate, and there were Blanks were obtained by adding TCA to the substrate
three replicates for each fish species. prior to incubation. The amino acid liberated was
measured following Moore and Stein (1948). Prote-
Inhibition assay ase activity was expressed as lg of glycine liberated
h-1 mg protein-1.
Inhibitory effect of tannin extracted from Pistia leaf
meal on the activity of principal digestive enzymes a-Amylase assay
were studied in vitro by adding graded levels of
extracted tannin (12.5–200 lg of tannin) to the test a-Amylase activity was determined following the
tubes containing enzyme extracts. Then, substrate and method described by Bernfeld (1955). Graded levels
PBS were added to the tubes and incubated at 37°C of extracted tannin were added to 1 ml of enzyme
for the optimal time period. Any minor change in pH extract followed by 1 ml of substrate (1% soluble
and volume of the reaction mixture due to the starch). The mixture was incubated at 37°C for
addition of tannin was corrected by adding PBS. A 20 min. Then, 2 ml of dinitrosalicylic acid reagent
concurrent control set without any tannin was main- (DNSA reagent, containing 1% dinitrosalicylic acid,
tained for each experimental set. Decrease in the 30% sodium potassium tartrate in 0.4 N NaOH
enzyme activity due to the addition of tannin was solution) was added to each tube and kept in a
expressed as percent reduction when compared to the boiling water bath for 5 min. Then, the tubes were
control set (100% activity). There were three repli- cooled and intensity of the color developed was read
cates for each of the experimental set. at 540 nm. Blanks were obtained by adding DNSA
reagent prior to incubation. Amylase activity was
Protease assay expressed as mg maltose liberated h-1 mg protein-1.

Protease activity was measured according to Anson Lipase assay

(1938) using bovine serum albumin (BSA) as
substrate. Graded levels of extracted tannin were Lipase activity was measured following the method
added to 0.1 ml of enzyme extract followed by described by Colowick and Kaplan (1955) using olive
0.1 ml of substrate (10 mg ml-1 BSA solution), oil as substrate. Briefly, graded levels of extracted
0.5 ml of phosphate buffer (pH 7.4, 0.1 M) and tannin (12.5–200 lg of tannin) were added to 0.5 ml
0.3 ml of distilled water. The reaction mixture was of enzyme extract followed by 2.5 ml of substrate
incubated for 1 h at 37°C. One milliliter of 10% (olive oil emulsion in 2% polyvinyl alcohol) and
trichloroacetic acid (TCA) was then added to stop the 0.5 ml of calcium chloride solution (110 mM). The
reaction and precipitate any remaining substrate. It reaction mixture was taken in 100-ml conical flask
was then centrifuged at 3,000 rpm for 10 min. To and incubated at 37°C for 1 h in a shaker incubator
2 ml of the supernatant, 1 ml of distilled water and (Lab. companion, SL-300R) with continuous shaking

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1174 Fish Physiol Biochem (2010) 36:1171–1180

(120 rpm). Then, 10 ml of acetone–ethanol mixture and C. mrigala were 14.85 (±0.15), 11.52 (±0.07)
(1:1) was added to stop the reaction. Few drops of and 10.32 (±0.13) lg of glycine liberated h-1 mg
phenolphthalein indicator were added, and fatty acid protein-1, respectively. There was a reduction of 9.65
liberated as a result of enzymatic action was titrated (±0.13)% to 88.28 (±0.68)% enzyme activity in
with 0.02 N NaOH solutions till the appearance of L. rohita when 12.5–200 lg of tannin was added.
faint pink color. One milliliter of 0.02 N NaOH is With similar doses of tannin, reduction in the
equivalent to 100 lM of free fatty acid. Blanks were protease activities ranged between 4.37 (±0.06)%
obtained using boiled enzyme. Lipase activity was to 43.96 (±0.09)% and 6.43 (±0.17)% to 82.53
expressed as l mole of fatty acid liberated h-1 mg (±0.99)% in C. catla and C. mrigala, respectively.
protein-1. Amylase activities in the control sets were 7.34
(±0.04), 8.43 (±0.06) and 6.73 (±0.07) mg maltose
Statistical analysis liberated h-1 mg protein-1 in L. rohita, C. catla and
C. mrigala, respectively. Reduction in the enzyme
The data obtained on the enzyme activity in presence activity due to the addition of tannin was reduced as
of the tannin were subjected to regression analysis, 3.22 (±0.07)% to 37.88 (±0.77)% in L. rohita, 3.12
using the concentration of the inhibitor as the (±0.03)% to 21.63 (±0.33)% in C. catla and 3.67
independent variable. Further, the data on the prote- (±0.14)% to 32.74 (±0.39)% in C. mrigala.
ase, amylase and lipase enzymes were applied for Activities of lipase in the control sets in L. rohita,
three-way ANOVA followed by Tukey’s test to infer C. catla and C. mrigala were recorded as 12.27
about the differences in the enzyme inhibition (±0.11), 9.17 (±0.05) and 10.82 (±0.06) lmol of
between the three fish species. All the statistical fatty acid liberated h-1 mg protein-1, respectively.
analyses were carried out following Zar (1999) using Reduction in the enzyme activities varied between
SPSS Ver10 (Kinnear and Gray 2000) software. 7.93 (±0.2)% to 87.07 (±0.59)% in L. rohita, 4.43
(±0.04)% to 41.35 (±0.4)% in C. catla and 6.41
(±0.03)% to 75.35 (±0.62)% in C. mrigala with the
Results addition of tannin.
The results of the three-way ANOVA (Table 4)
The tannin content in powdered Pistia was revealed significant differences in enzyme activity
26.67 mg g-1 of leaf meal. The concentration of due to tannin in terms of dose, type of enzyme and
tannin in the tannin extract was 0.89 mg mL-1. The the species. The post hoc Tukey test revealed
results of the experiment revealed that the Pistia significant differences in protease, amylase and lipase
tannin (PT) significantly inhibit/lower the activities activity between the three fish species (Between
of the digestive enzymes from three IMCs in a dose L. rohita and C. catla: 1.09; between L. rohita and
dependant manner (Fig. 1) as evident from the C. mrigala: 0.51; between C. catla and C. mrigala:
regression equations (Table 2). In all the instances 1.61; for all values P \ 0.001).
irrespective of the carp species, the enzyme activity
was noted to decrease with the increase in the tannin
concentrations. However, variation in the reduction Discussions
of enzyme activities was noticed between the three
fish species, as well as the enzymes studied. Among Chemical compounds that render plant tissues unpal-
the fish species, digestive enzymes from L. rohita atable are known to be widely distributed in the plant
were found to be the most sensitive to the PT kingdom (Liener 1989; Makkar 1993; Hagerman et al.
followed by C. mrigala, whereas enzymes from 1997). Tannins are one such group of phenolic
C. catla were comparatively least affected. The compounds which have received a lot of attention
activities of protease and lipase were more affected with respect to their possible nutritional and physio-
compared to the amylase. logical interference. The nutritional value and possi-
Percent inhibition of the enzyme activities are bilities of utilization of aquatic weeds have been
presented in Table 3. Protease activities in the evaluated in some of the literatures in the last decade
control sets (without tannin) in L. rohita, C. catla (Edwards et al. 1985; Ray and Das 1992, 1994, 1995;

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Fish Physiol Biochem (2010) 36:1171–1180 1175

Fig. 1 The activity of the intestinal protease (lg glycine liberated mg-1 protein h-1), amylase (mg maltose liberated mg-1 protein
h-1) and lipase (lmol fatty acid liberated mg-1 protein h-1) from the Indian major carps under different doses of tannins (lg)

Hasan et al. 1990; Patra and Ray 1988; Patra et al. revealed tannin concentration as high as
1999, 2002). In view of accessing effectiveness of 26.67 mg g-1 in the Pistia leaf meal. Such com-
aquatic weeds in carp diet formulation, quantification pounds or antinutritional factors are believed to be the
of tannin in the dried Pistia leaf meal has been defense weapon of plants against the herbivores
performed in the present investigation. Previous study (Becker and Makkar 1999). While studying the
by Mandal and Ghosh (2009a) depicted tannin enzyme inhibitory effect of the Pistia tannin in the
concentration in some natural and potential fish food present study, it has been revealed that the digestive
items/ingredients of plant origin that ranged between enzymes from L. rohita were most sensitive to the
5.38 (coconut oil cake) to 34.3 (phytoplankton) Pistia tannin (PT) whereas enzyme from C. catla were
mg g-1 dry weight. Result of the present study comparatively least affected. Herbivory is more

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1176 Fish Physiol Biochem (2010) 36:1171–1180

Table 2 Regression equations of the activities of enzymes from the Indian major carps against graded levels of tannin
Enzymes Fish species Regression equation
(y = enzyme activity, x = dose of tannin)

Protease L. rohita y = 13.36 - 0.064x; F = 181.98, df = 1,19; p \ 0.001; R2 = 0.905

C. catla y = 11.37 - 0.023x; F = 834.63, df = 1,19; p \ 0.001; R2 = 0.978
C. mrigala y = 10.12 - 0.04x; F = 848.1, df = 1,19; p \ 0.001; R2 = 0.978
Amylase L. rohita y = 7.08 - 0.013x; F = 254.35, df = 1,19; p \ 0.001; R2 = 0.93
C. catla y = 8.3 - 0.009x; F = 614.45, df = 1,19; p \ 0.001; R2 = 0.97
C. mrigala y = 6.58 - 0.01x; F = 304.04, df = 1,19; p \ 0.001; R2 = 0.941
Lipase L. rohita y = 11.98 - 0.05x; F = 2687.66, df = 1,19; p \ 0.001; R2 = 0.993
C. catla y = 8.84 - 0.02x; F = 348.86, df = 1,19; p \ 0.001; R2 = 0.948
C. mrigala y = 10.49 - 0.04x; F = 2099.89, df = 1,19; p \ 0.001; R2 = 0.991

common in the L. rohita than C. mrigala; on the other inhibition of digestive enzymes in carps must be from
hand, C. catla feeds on zooplanktons (Jhingran 1997; their exposure to the hydrolysable tannin (or, tannic
Mohanty 2003). Most of the previous studies reported acid), indicating its potential to interfere with the
L. rohita to be herbivore (Alikunhi 1958; Khan and digestive process.
Siddiqui 1973); however, some of the recent inves- Commonly used ingredients in formulated fish
tigations have focused on omnivorous feeding apti- feed like mustard oil cake and sun flower oil cake
tude of this species by analyzing feed intake/ contain tannin as high as 21.25 mg and
preference (Mohanty 2003; Rahman et al. 2006) or 26.25 mg g-1 dry weight, respectively (Mandal and
symbiotic gut microbial community (Ghosh et al. Ghosh 2009a). It may be assumed that if fish of 100 g
2002). Therefore, maximum inhibition of digestive biomass is fed at the rate of 3% of the body weight
enzymes from L. rohita by plant tannin and appear- daily with a formulated diet having only 10% of these
ance of omnivorous feeding aptitude may be viewed ingredients, the fish will be exposed to more than
as co-evolution where tannin in the leaf meal appears 6 mg of tannin daily. Natural feeding on phytoplank-
to play as a defense compound. To reduce damage ton tends to increase this value. In the present study,
caused by herbivory, plants have evolved a broad concentration of tannin used (12.5–200 lg) are much
array of defenses, majority of which are chemicals. lower than this presumptive value. The results of the
Herbivores prefer to avoid ingesting tannin if alternate present study clearly indicated that tannin, even in
foods are available (Wynne-Edwards 2001). Thus, very low concentration, can inhibit or lower the
defenses and counter defenses lay down the step for activities of protease, amylase and lipase in all the
the co-evolution of plants and herbivores. three carp species studied, which is in agreement with
Hydrolysable tannins are easily degraded in bio- the study made by Maitra and Ray (2003) in L. rohita
logical systems by non-specific esterase, and the fingerlings. In a study based on growth and hemato-
hydrolyzed products entering into the blood may immunological parameters, Prusty et al. (2007)
cause organ toxicity (particularly in liver and kidney) opined that tannic acid in feed (up to 2%) may not
once the level in blood rises beyond the detoxification be detrimental for Labeo rohita fingerlings. However,
capability of these organs (Garg et al. 1992; Muller- they did not address enzyme activities in the short-
Harvey and McAllan Mueller-Harvey and McAllan term feeding trial of 60 days. Similarly, Becker and
1992). In contrast, the condensed tannins are complex Makkar (1999) assumed that tannic acid may not
and larger molecules which are not hydrolyzed in the affect nutrient availability and enzyme activity in
biological system, indicating their least effect in fish gastrointestinal tract of carp. In contradiction to these
(Makkar et al. 1995). Condensed tannin did not observations, in vitro studies made in the present
appear to bind to proteins and other nutrients and experiment clearly indicated inhibition of the diges-
decrease their bioavailability in the carp intestine tive enzyme activities in carps by the tannic acid
(Becker and Makkar 1999). In the present study, extracted from the Pistia. Although experiment

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Fish Physiol Biochem (2010) 36:1171–1180 1177

6.41 ± 0.03
12.17 ± 0.16

59.09 ± 0.15
25.79 ± 0.16

75.35 ± 0.62
43.1 ± 0.67
Table 4 Results of the three-way factorial ANOVA for the

C. mrigala
effect of tannin on the enzyme activities in the Indian major
carps (F-values are significant at P \ 0.001)
Table 3 Percent reduction of in vitro activities of the intestinal enzymes from the three Indian major carps by graded levels of tannin extracted from Pistia leaf meal

Source Sum of squares df Mean square F-value

4.43 ± 0.04 DOSE (D) 631.45 5 126.29 9,222.37

37.28 ± 0.75
17.62 ± 0.36
29.87 ± 1.04
8.9 ± 0.1

41.35 ± 0.4
ENZYME (E) 51.07 2 25.54 1,864.87
C. catla

FISH SP. (F) 72.76 2 36.38 2,656.61

D9E 142.28 10 14.23 1,038.99
D9F 81.24 10 8.12 593.26
12.95 ± 0.12
24.69 ± 0.13

87.07 ± 0.59
49.28 ± 0.67
62.81 ± 0.41
E9F 36.78 4 9.19 671.55
7.93 ± 0.2

D9E9F 43.79 20 2.19 159.91

L. rohita
Lipase

Error 1.48 108 0.01

Total 1,060.86 161
3.67 ± 0.14
5.75 ± 0.14
14.42 ± 0.29

32.74 ± 0.39
17.29 ± 0.15
21.84 ± 0.53

conducted with commercial tannin may not corre-

C. mrigala

spond with the tannins of the feed components

(Becker and Makkar 1999). Tannins may reduce
macro nutrient utilization by forming tannin–protein
complexes with various digestive enzymes preclud-
3.12 ± 0.03
3.68 ± 0.08
7.79 ± 0.16

21.63 ± 0.33
12.65 ± 0.23
16.81 ± 0.3

ing the formation of the product absorbable by the

C. catla

small intestine (Carmona et al. 1996). In addition, it

% Reduction in enzyme activity (control, without tannin at 100% activity)

may inhibit transport systems concerning carbohy-

drate assimilation such as glucosidase/maltase
(Bjorck and Nyman 1987), sucrase (Welsch et al.
3.22 ± 0.07
9.31 ± 0.23
18.53 ± 0.31

37.88 ± 0.77
22.35 ± 0.37
26.52 ± 0.28

1989) and the intestinal sodium-dependent glucose

L. rohita
Amylase

uptake system (Karasov et al. 1992). Bean tannins

were shown to strongly inhibit pancreatic trypsin,
chymotrypsin and a-amylase (Sing 1984; Carmona
6.43 ± 0.17
10.66 ± 0.19
28.2 ± 0.19

82.53 ± 0.99
38.61 ± 0.48
54.12 ± 0.42

et al. 1991).
C. mrigala

Tannins are considered as plant secondary metab-

olites which are distinguished from other polypheno-
lic compounds by their ability to precipitate proteins
(Silanikove et al. 2001). It has been postulated that
4.37 ± 0.06
6.74 ± 0.14
13.23 ± 0.21

43.96 ± 0.09
20.75 ± 0.11
27.47 ± 0.27

tannins interfere with protein and dry matter digest-

ibility by inhibiting protease and also forming
Values are means ± SE of three determinations
C. catla

indigestible complexes with dietary protein (Krog-

dahl 1989). However, little is known about the effect
of tannin on fish (Makkar and Becker 1998). Vohra
9.65 ± 0.13
17.87 ± 0.35
47.17 ± 0.51

88.28 ± 0.68
67.35 ± 0.56
71.42 ± 0.6

et al. (1966) reported that tannins caused growth

L. rohita
Protease

depression in chickens at levels as low as 0.5% of the

diet. Similarly, Hossain and Jauncey (1989) observed
poor growth response in common carp (Cyprinus
carpio) fed diets containing 0.57 and 1.14% tannins.
Tannin added (lg)

In addition, high levels of tannins in feed have been

shown to have adverse effect on herbivorous and
omnivorous fish (Al-Owafeir 1999; Becker and
Makkar 1999; Olvera et al. 1988). The extents of
12.5

200
100
150
25
50

these growth inhibitory effects varied in different fish

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1178 Fish Physiol Biochem (2010) 36:1171–1180

species studied and also plant material used in the fish (FIST programme), New Delhi, India for providing research
feed formulation. Present study also revealed varied facilities. The authors are obliged to Dr. G. Aditya, Department
of Zoology, The University of Burdwan for rendering help in
degrees of tannin-induced inhibition of digestive statistical analyses of data. The first author is grateful to The
enzymes from different carp species studied. In University of Burdwan, Burdwan for awarding the university
agreement with the reports discussed above, it may fellowship.
be assumed from the results of the present study that
tannin may affect feed utilization efficiency and References
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