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Bioresource Technology 85 (2002) 1724

Duckweed (Lemna polyrhiza) leaf meal as a source of feedstu


in formulated diets for rohu (Labeo rohita Ham.) ngerlings
after fermentation with a sh intestinal bacterium
A. Bairagi a, K. Sarkar Ghosh a, S.K. Sen b, A.K. Ray

a,*

a
b

Fisheries Laboratory, Department of Zoology, Visva-Bharati University, Santiniketan 731 235, West Bengal, India
Microbiology Laboratory, Department of Botany, Visva-Bharati University, Santiniketan 731 235, West Bengal, India
Received 29 December 2001; received in revised form 20 February 2002; accepted 12 March 2002

Abstract
Eight isonitrogenous (35% crude protein approximately) and isocaloric (4.2 kcal g1 approximately) diets were formulated including raw and fermented duckweed (Lemna polyrhiza) leaf meal at 10%, 20%, 30% and 40% levels. A particular bacterial strain
(Bacillus sp.) isolated from carp (Cyprinus carpio) intestine and having extracellular amylolytic, cellulolytic, proteolytic and lipolytic
activities was used for leaf meal fermentation for 15 days at 37 C. The bre content of leaf meal reduced from 11.0% to 7.5% and
the antinutritional factors, tannin and phytic acid, were reduced from 1.0% to 0.02% and 1.23% to 0.09%, respectively after fermentation. However, the available reducing sugars, free amino acids and fatty acids increased in the fermented leaf meal. The
response of rohu, Labeo rohita, ngerlings fed the experimental diets for 80 days was compared with sh fed a sh meal based
reference diet. On the basis of growth response, food conversion ratio and protein eciency ratio, 30% fermented Lemna leaf meal
incorporated in the diet resulted in the best performance of rohu ngerlings. In general, growth and feed utilization eciencies of
sh fed fermented leaf meal containing diets were superior to those fed diets containing raw leaf meal. The apparent protein digestibility (APD) decreased with increasing levels of leaf meal irrespective of treatment. The APD for raw leaf meal was lower at all
levels of inclusion in comparison to those for the fermented meals. The highest carcass protein and lipid deposition was recorded in
sh fed the diet containing 30% fermented leaf meal. The results showed that fermented Lemna leaf meal can be incorporated into
carp diets up to 30% level compared to 10% level of raw meal. 2002 Elsevier Science Ltd. All rights reserved.
Keywords: Lemna polyrhiza; Intestinal bacteria; Fermentation; Diets; Growth performance; Labeo rohita ngerlings

1. Introduction
Considering the importance of nutritionally balanced
and cost-eective articial diets for sh, there is an increasing research eort to evaluate the nutritive value
of dierent non-conventional feed resources, including terrestrial and aquatic macrophytes (Edwards et al.,
1985; Wee and Wang, 1987; Patra and Ray, 1988; Ray
and Das, 1992, 1993, 1995; Mondal and Ray, 1999).
Aquatic and terrestrial macrophytes have been used as
supplementary feeds in sh farming since the early times
of freshwater sh culture (Bardach et al., 1972) and still
play an important role as sh feed in extensive culture systems (Edwards, 1987). The aquatic weeds have
been shown to contain substantial amounts of protein

Corresponding author.

and minerals (Ray and Das, 1994). These weeds, which


otherwise remain unutilized, and often make the water
body unsuitable for sh culture, may be converted into
valuable sh esh through their incorporation as a feedstu in carp diets. However, the presence of antinutritional factors within plant feedstus restricts their use in
animal feeds (Tacon, 1990). Processing plant materials
through a simple and cheap method like fermentation
might considerably decrease the antinutritional factors
and crude bre content thereby increasing the plants
nutritional values. The ecacy of various plant sources
for partial or complete replacement of sh meal in
aquafeeds has been investigated by a number of workers
(Atack and Matty, 1979; Viola et al., 1982; Hossain and
Jauncy, 1989; Tacon, 1993; Ray and Das, 1995; Mondal
and Ray, 1999). The Indian major carp, L. rohita is
primarily a herbivorous to omnivorous species and
prefers to feed on plant materials (Talwar and Jhingran,

0960-8524/02/$ - see front matter 2002 Elsevier Science Ltd. All rights reserved.
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A. Bairagi et al. / Bioresource Technology 85 (2002) 1724

1991). Lemna polyrhiza, commonly known as duckweed,


grows luxuriantly in freshwater bodies in the tropical
and subtropical areas including India and Bangladesh
(Majid, 1986). This aquatic weed grows vigorously
during the rainy season and generally disappears at the
end of winter. These are free-oating aquatic plants
which are not accepted by cattle and Indian major
carps as feed in fresh condition. This aquatic weed has
been shown to contain substantial amounts of protein and minerals (Ray and Das, 1994). These weeds,
which otherwise remain unutilized and often makes
the water body unsuitable for sh culture, may be converted into valuable sh esh through their incorporation as a feedstu in carp diets. The use of fermented
aquatic weed as one of the ingredients in carp diets will
denitely bring down the cost of sh production. In the
present study, Lemna polyrhiza was fermented by a
cellulose-degrading sh gut bacterium, Bacillus sp.
and an experiment was designed to evaluate the possible utilization of fermented Lemna meal as a partial
substitute of dietary sh meal protein in diets for the
Indian major carp, rohu, L. rohita (Hamilton) ngerlings.

2. Methods
2.1. Isolation and characterization of sh intestinal
bacteria
The bacterial strain used for fermentation of Lemna
leaf was isolated from the intestine of common carp,
Cyprinus carpio, on sterilized carboxymethylcellulose
(CMC)-agar medium (pH 7) containing (g l1 ), CMC,
10; KH2 PO4 , 4; Na2 HPO4 , 4; MgSO4  7H2 O, 0.2; CaCl2 ,
0.001; FeSO4  7H2 O, 0.004; Agar, 15. CMC-ase producing isolates were identied on CMC plates after
ooding the plates with 5 ml of Congo red dye prepared
in 0.7% agarose (Seakem HGT agarose) essentially
according to the method of Teather and Wood (1982).
The bacterial isolate was further screened for the production of extracellular amylase (Bernfeld, 1955), cellulase (Denison and Koehn, 1977), protease (Walter, 1984)
and lipase (Colowick and Kaplan, 1955). The strain was
identied and characterized by carrying out the tests
described in the Mannual of Microbiological Methods
(Society of American Bacteriologists, 1957).
2.2. Preparation of bacterial seed culture
The selected bacterium was grown in shake bottles in
4% tryptone soya broth (Hi-media, India) for seed culture. After 24 h of growth at 37 C, an average viable
count was about 107 cells/ml of broth. This was used as
bacterial seed for Lemna leaf fermentation.

2.3. Fermentation of Lemna leaf


The leaves of Lemna polyrhiza were sundried. The
dried leaves were nely ground and passed through a
ne meshed sieve to ensure homogeneity. A portion of
sieved Lemna leaf was moistened with 50% w/v liquid
basal medium containing (g l1 ): KH2 PO4 , 4; Na2 HPO4 ,
4; MgSO4  7H2 O, 0.2; CaCl2 , 0.001; FeSO4  7H2 O,
0.004 and autoclaved for sterilization. The sterilized leaf
meal was fermented with Bacillus culture at the rate of
108 bacterial cells/g of dried leaf for 15 days at 37 C in
an incubator.
2.4. Diet preparation
For formulation of test diets, both fermented and
unfermented or raw leaf meals were used. Two sets of
experimental diets were formulated using either raw
(diets D1 to D4) or fermented (diets D5 to D8) Lemna
meal at 10%, 20%, 30% or 40% levels (Table 2). A diet
containing sh meal as the main protein source was used
as the reference diet (RD). To each of the formulated
diets, 1% chromic oxide was added as digestibility
marker. All the diets were prepared in pelleted form
using 0.5% carboxymethylcellulose as a binder. The
pellets were sun dried for a few days and crumbled prior
to feeding.
2.5. Experimental design
The experiment was conducted in ow-through 90l
circular bre-glass tanks. Rohu ngerlings, obtained
from a local sh dealer, were acclimatized to the laboratory conditions for 15 days and fed with a mixture of
ricebran and mustard oil cake. The ngerlings (mean
weight 6:4  0:31 g) were randomly distributed at the
rate of 15 sh per tank with three replicates for each
treatment. Each experimental tank was supplied with
unchlorinated water from a deep tube well with continuous aeration. All the sh were fed once daily at 1000
h at a xed feeding rate of 3% body weight per day for
80 days. The quantity of feed given was readjusted every
15th day after weighing the sh. To determine the feed
consumption, any left-over feed was collected 6 h after
each feeding and weighed after oven drying. The faecal
samples were collected everyday in the morning by siphoning at 17 h after removal of the uneaten feed, and
following the immediate pipetting method outlined by
Spyridakis et al. (1989), from three replicates of each
dietary treatment. The faeces naturally released by the
sh could be easily detected and were immediately removed from the water with a glass canula. At the termination of the 80 day experiment the sh were weighed
and analyzed for carcass composition.
The water quality parameters from each tank were
monitored each week throughout the experimental

A. Bairagi et al. / Bioresource Technology 85 (2002) 1724

period. The ranges of water quality parameters were:


temperature, 2930 C; pH 77.5; dissolved oxygen, 4.5
5.3 mg l1 and alkalinity 160175 mg l1 .
2.6. Chemical analyses and data collection
Proximate analysis of feed ingredients, experimental diets and faecal samples was performed following
the AOAC (1990) procedures as follows: moisture was
determined by oven drying at 105 C for 24 h; crude
protein (Nitrogen  6:25) by micro Kjeldahl digestion
and distillation after acid digestion using a Kjeltec 1026
Distilling Unit together with a Tecator Digestion System
(Tecator, Sweden); lipid was determined by extracting
the residue with 4060 C petroleum ether for 78 h in a
Soxhlet apparatus; crude bre was determined as loss on
ignition of dried lipid-free residues after digestion with
1.25% H2 SO4 and 1.25% NaOH; and ash was determined by ignition at 550 C in a Mue furnace to constant weight. Nitrogen-free extract (NFE) was computed
by taking the sum of values for crude protein, crude
lipid, crude bre and moisture and subtracting this
from 100 (Maynard et al., 1979). Five sh from each
aquarium were sampled at the termination of the feeding experiment, and the whole body was analysed for
moisture, crude protein, crude lipid and ash following
the aforementioned methods. Chromic oxide in the diets
and faecal samples was estimated following the method
of Bolin et al. (1952). Tannin content in both fermented and raw Lemna meal was determined using Folin
Denis reagent (Schanderi, 1970). Phytic acid content was
determined according to Wheeler and Ferrel (1971).
Estimation of total free amino acids was conducted according to Moore and Stein (1948) using ninhydrin reagent dissolved in methyl cellosolve. Total free fatty
acids in raw and fermented leaf meal were estimated
following the method described by Cox and Pearson
(1962). The water quality parameters were monitored
following the methods outlined by APHA (1985). Ap-

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parent protein digestibility (APD, %), specic growth


rate (SGR, % day1 ), feed conversion ratio (FCR), protein eciency ratio (PER) and apparent net protein
utilization (ANPU, %) were calculated using standard
methods (Steens, 1989).
2.7. Statistical analysis
Statistical analysis of data was performed by analysis
of variance (ANOVA) using Microsoft software Statistica followed by Duncans multiple range test (Duncan,
1955).

3. Results
The proximate compositions of feed ingredients and
experimental diets are presented in Tables 1 and 2, respectively. Fermentation of Lemna leaf meal resulted in a
signicant decrease in the levels of crude bre and antinutritional factors, tannin and phytic acid, whereas,
there was increase in the levels of free amino acids and
fatty acids. A comparison of the proximate composition
of the reference diet, and Lemna leaf meal incorporated
diets indicated that the crude bre level in the control diet
was 11.80%, whereas, it ranged from 7.87.9% in diets
containing raw Lemna leaf meal, and reduced to 3.8
5.8% in diets containing fermented Lemna leaf meal. In
diets containing fermented Lemna leaf meal the tannin
and phytic acid contents were below detection limit.
The growth performance and feed utilization of L.
rohita ngerlings in terms of percentage weight gain,
SGR, FCR, PER, ANPU and APD are presented in
Table 3. The average weight of the sh increased in all
the dietary treatments, in comparison to the reference
diet excepting in diet D4, which contained 40% raw
Lemna leaf meal. The highest attainment in sh body
weight, average percentage live weight gain and SGR
were recorded in the group of sh reared on diet D7

Table 1
Proximate compositions of feed ingredients (% dry matter basis)
Nutrients

Fish meal

Mustard oilcake

Rice bran

Raw Lemna leaf meal

Fermented Lemna leaf meal

Moisture
Dry matter
Crude protein
Crude lipid
Ash
Crude bre
NFE
Gross energy
(kcal g1 )
Tannin
Phytic acid
Total free amino
acid
Total free fatty acid

2.26
97.04
58.50
8.91
11.50
3.93
14.20
4.19

14.00
86.00
35.95
7.00
8.37
5.53
29.17
4.11

4.45
95.55
13.00
5.14
21.41
25.50
30.50
3.52

32.50
67.50
18.60
1.50
2.50
11.00
83.90
5.04

11.38
1.00

7.50

1.00
1.23
0.32

0.02
0.09
0.95

2.0

6.0

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A. Bairagi et al. / Bioresource Technology 85 (2002) 1724

Table 2
Ingredient composition (% dry weight) and proximate analyses of experimental diets (on dry matter basis)
RD

Diets with Lemna leaf meal


Raw

Ingredients
Fish meal
Mustard oilcake
Rice bran
Lemna meal
Premixa
Chromic oxide
Proximate composition
Dry matter
Crude protein
Crude lipid
Ash
Crude bre
NFEc
Gross energy
(kcal g1 )
Tannins
Phytic acid
Chromic oxide

Fermented

D1

D2

D3

D4

D5

D6

D7

D8

40.0
23.0
35.0

1.0
1.0

36.0
25.0
27.0
10.0
1.0
1.0

34.0
24.0
20.0
20.0
1.0
1.0

32.0
24.0
12.0
30.0
1.0
1.0

30.0
28.0

40.0
1.0
1.0

36.0
25.0
27.0
10.0
1.0
1.0

34.0
24.0
20.0
20.0
1.0
1.0

32.0
24.0
12.0
30.0
1.0
1.0

30.0
28.0

40.0
1.0
1.0

(%)b
98.0
35.95
8.50
10.0
11.80
31.50
4.6

94.0
35.41
8.50
12.0
7.80
30.89
4.2

98.0
34.93
8.50
14.0
7.90
32.83
4.3

98.0
34.88
8.0
14.0
7.80
32.80
4.4

98.0
35.26
8.50
20.0
7.90
26.20
4.2

96.0
34.68
5.0
12.0
3.80
50.70
4.1

98.0
34.58
5.5
16.0
4.00
51.50
3.9

94.0
33.29
6.50
16.0
4.80
38.72
3.9

96.0
32.97
6.0
20.0
5.80
35.91
3.9

ND
ND
0.97

0.1
0.12
0.92

0.2
0.24
0.95

0.3
0.36
0.97

0.4
0.48
1.0

ND
ND
1.01

ND
ND
1.02

ND
ND
1.01

ND
ND
1.03

ND Not detectable.
a
Vitamin and mineral mixture (Vitaminetes Forte, Roche Products Ltd, 24/28 Pt. M.M. Malaviya Road, Mumbai 400 034, India).
b
Number of samples for each determination 3.
c
Nitrogen-free extract.

(30% fermented Lemna meal incorporation). Fish reared


on diet D6 (20% fermented Lemna meal) also showed
good performance in terms of live weight gain (%) and
SGR. PER was highest in sh fed diet D7 which was
signicantly dierent (P < 0:05) from those reared with
other diets. PER value was lowest with the reference diet
(without Lemna meal substitution). The FCR value was
lowest for sh fed diet D7 and highest for the reference
diet.
ANPU was highest in sh fed diet D7 and lowest with
diet D2. Apparent protein digestibilities (APD) for all
diets were, however, high, ranging from 82.30% to
94.43%. Rohu ngerlings fed diet D5 showed the highest
APD value (94.43%). Poor protein digestibility was recorded in sh fed diet D4, containing raw Lemna leaf
meal at 40% level of incorporation, and also with diet
D8, containing fermented Lemna leaf meal at the same
level of incorporation.
The carcass composition of experimental sh before
and after the experiment is given in Table 4. Rohu ngerlings fed diet D8 had a slightly higher percentage of
moisture than in sh fed other experimental diets. The
deposition of protein in the carcass of experimental sh
increased over the initial value in all the dietary treatments. Although all the sh were fed isonitrogenous
diets, the greatest accumulation of carcass protein was
recorded in the group of sh reared on diet D7, containing 30% fermented Lemna leaf meal. Highest tissue

lipid accumulation occurred in sh fed diet D7. The


carcass lipid content varied signicantly (P < 0:05)
among dierent dietary treatments. The ash content of
sh carcass was highest (P < 0:05) in sh diet D6.

4. Discussion
It is evident from the present study that fermented
duckweed meal can be utilized as a feed ingredient in
the diets for the rohu, L. rohita ngerlings eectively up
to 30% level of incorporation without compromising
growth. Growth performance, PER and FCR of rohu
ngerlings were better with 40% fermented Lemna leaf
meal incorporated diet than in the reference diet. Das
and Ray (1989) demonstrated the possibility of incorporation of dried Lemna polyrhiza as a feed ingredient
for L. rohita ngerlings, and recorded higher carbohydrate digestibility in relation to that of the control diet
although the SGR and percent weight gain in sh fed
Lemna meal were slightly lower than with the control
diet. Shireman and Smith (1983) considered duckweed
(Lemna sp.) as a highly nutritious vegetable food for
grass carp because of its tenderness and high protein
content compared to other aquatic weeds. In the present
investigation, the protein content of Lemna polyrhiza
was estimated to be 18.60%. However, the presence of
antinutritional factors limits direct use of the weed as a

Table 3
Growth, feed utilization eciencies and APD in L. rohita ngerling fed experimental diets for 80 days. Data are mean values  SE (n 5)
Diets with Lemna leaf meal diet

RD

D1

D2

D3

D4

D5

D6

D7

D8

Initial weight (g)


Final weight (g)
Weight gain (%)
Feed intake1 (g/
100 g BW of sh/
day)

SGR (% day1 )

FCR

PER

ANPU (%)

APD (%)

6.4  0.31
11.02  0.29a
72.18  0.12b
1.65

6.4  0.32
11.22  0.31a
75.31  0.16e
1.43

6.4  0.31
11.20  0.33a
75.00  0.14d
1.48

6.4  0.34
11.18  0.28a
74.78  0.10c
1.52

6.4  0.33
10.80  0.34a
68.75  0.18a
1.52

6.4  0.33
11.72  0.32b
83.12  0.11g
1.54

6.4  0.32
11.89  0.30b
85.78  0.18h
1.47

6.4  0.31
11.93  0.31b
86.50  0.16i
1.48

6.4  0.34
11.54  0.33b
80.31  0.15j
1.47

0.905  0.03a
3.17  0.10c
0.87  0.06a
11.47  0.50c
89.46  4.25b

0.935  0.03b
2.68  0.19ab
1.05  0.05b
4.62  0.21b
86.78  4.24a

0.931  0.02b
2.79  0.06b
1.03  0.06b
2.58  0.12a
90.30  4.22b

0.931  0.04b
2.87  0.15b
1.00  0.04a
37.60  1.08f
86.20  4.22a

0.876  0.02a
3.01  0.13c
0.94  0.04a
31.89  1.50e
82.30  3.21a

1.00  0.03c
2.73  0.09ab
1.05  0.05b
26.34  1.30d
94.43  4.24b

1.031  0.02c
2.56  0.11a
1.13  0.05b
72.87  3.16g
93.90  4.21b

1.038  0.03c
2.55  0.12a
1.18  0.06b
98.71  4.52h
90.20  4.19b

0.981  0.04b
2.66  0.15ab
1.14  0.06b
5.58  0.25b
85.60  4.06a

Means with same superscripts in the same row were not signicantly dierent (P < 0:05).
BW: body weight.

SGR ln final weight  ln initial weight=Days on trial  100;  PER Wet weight gain of the fish=Protein consumed;  FCR Dry weight of the feed given=Increase in wet weight of
fish;  ANPU Net increase in carcass protein=Amount of protein consumed  100;  APD 100  100  %Cr2 O3 in diet=%Cr2 O3 in faeces  %protein in faeces=%protein in diet.
1
Statistical analysis was not possible as determinations were performed on pooled samples.

Table 4
Proximate carcass compositions (% wet weight) of the experimental sh at the start and end of the 80 day feeding experiment. Data are mean values SE (n 5)
Carcass composition

Initial

RD

Diets
D1

D2

D3

D4

D5

D6

D7

D8

Moisture
Crude protein
Crude lipid
Ash

76.31
12.90
2.26
4.96

75.27  0.17c
13.50  0.58a
2.66  0.04c
3.23  0.06a

75.19  0.13c
13.11  0.40a
2.16  0.08b
3.40  0.09ab

74.80  0.15b
13.02  0.48a
2.03  0.07a
3.68  0.11b

76.06  0.15ef
14.69  0.50b
1.83  0.12a
3.49  0.20b

75.60  0.09d
14.38  0.52b
1.83  0.18a
3.19  0.26a

75.86  0.10e
14.22  0.45b
2.23  0.11b
4.20  0.25c

73.23  0.16a
16.42  0.60c
2.30  0.13b
5.03  0.09d

75.93  0.13e
17.52  0.65d
2.70  0.09c
5.00  0.14d

76.18  0.14f
13.15  0.42a
2.16  0.15b
4.24  0.09c

A. Bairagi et al. / Bioresource Technology 85 (2002) 1724

Mean values

Means with same superscripts in the same row were not signicantly dierent (P < 0:05).

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A. Bairagi et al. / Bioresource Technology 85 (2002) 1724

dietary ingredient. Dietary tannin as low as 0.5% causes


growth depression in chickens (Vohra et al., 1996) and
there are also reports on the toxicity of tannin to sh
(Hossain and Jauncy, 1989; Krogdahl, 1989; Mukhopadhayay and Ray, 1999). In the present study, tannin
and phytic acid could not be detected in fermented
Lemna leaf meal incorporated diets. Fibre contents of
plant ingredients could also be responsible for their poor
digestibility (De Silva et al., 1990) apart from the presence of some antinutritional factors (Hastings, 1964;
Atack et al., 1979; Jackson et al., 1982). High levels of
bre in the diet are known to retard the growth of sh
(NRC-NAS, 1977; Edwards et al., 1985). A possible
reason for poor performance of the rohu ngerlings on
the reference diet may have been its relatively high level
of crude bre. Fish do not possess the enzyme cellulase
to hydrolyze cellulose which is the main ingredient of
plant cell walls and the possible presence of a persistent
population of cellulolytic microora in sh gastrointestinal tract is a subject of much controversy (Stickney
and Shumway, 1974; Prejs and Blaszczyk, 1977; Lindsay
and Harris, 1980; Lesel et al., 1986; Luczkovich and
Stellwag, 1993; Saha and Ray, 1998). In our previous
study on isolation of cellulolytic gut microora from
Indian major carps and Chinese carps we isolated certain carboxymethylcellulaseproducing strains (Bairagi
et al., 2000), and in the present study, a Bacillus sp. from
the common carp, Cyprinus carpio, was used to ferment
Lemna leaf meal in vitro. During fermentation, nutrient
losses may occur as a result of leaching, destruction by
light, heat or oxygen or microbial utilization (Jones,
1975). Nevertheless, loss of nutrients during fermentation is commonly negligible and there may be an increase in the nutrient level through microbial synthesis
(Wee, 1991). The protein utilization eciency of sh fed
fermented Lemna leaf meal incorporated diets was signicantly better than those fed raw leaf meal. Edwards
(1980) reported better growth of tilapia, Oreochromis
mossambicus, fed diets containing composted water hyacinth than with conventional tilapia feed, although the
latter contained more protein. Ray and Das (1992) also
indicated the possibilities of incorporation of composted
Salvinia cuculata in supplementary diets for the Indian
major carp, L. rohita substituting the conventional diet
up to 20% level. In the present study, the best performance of sh in terms of percentage weight gain, SGR,
FCR and PER was observed in sh fed a diet containing
30% fermented Lemna leaf meal which was followed by
the diet containing 20% fermented Lemna leaf meal.
Fish fed diet with 40% raw Lemna leaf meal gave the
poorest performance in terms of percentage weight gain
and SGR. Since all the diets were isonitrogenous, the
reduced growth of the sh fed diets containing higher
levels of raw Lemna leaf meal appeared to be due to
increasing bre content and antinutritional factors in the
diets.

In the present study, a progressive decline in APD


with increasing level of raw Lemna leaf meal was observed. A declining trend in APD values has also been
reported with higher levels of inclusions of raw mustard,
linseed, sesame seed raw copra meals in carp diets.
(Hossain and Jauncy, 1989; Hasan et al., 1991; Mukhopadhayay and Ray, 1999). The presence of antinutritional factors may inuence the digestibility of various
nutrients in the diet and give erroneous results (Lall,
1991). The lower APD values for the diets incorporating
raw Lemna meal in comparison to those containing
fermented Lemna meal may be due to the presence of
antinutritional factors in the raw Lemna meal.
The carcass protein and lipid contents were found to
increase with increasing level of incorporation of fermented Lemna leaf meal up to the 30% level, but declined in sh fed diet D8, containing 40% fermented leaf
meal.

5. Conclusion
It was concluded from the present study that fermented Lemna leaf meal can be recommended as a dietary ingredient in diets of L. rohita ngerlings up to
30% incorporation level without any adverse eect on
growth of the sh. The growth and feed utilization efciencies of sh fed diets incorporating fermented
Lemna leaf meal were better than those fed raw Lemna
leaf meal at the same level of inclusion.

Acknowledgements
We are grateful to the Indian Council of Agricultural
Research, New Delhi (project F. no. 4 (11)/98-ASR-I)
and University Grants Commission, New Delhi (DSA
Programme to the Department of Zoology) for nancial
support.

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