Plant Archives Volume 20 No. 2, 2020 pp.

5177-5185 e-ISSN:2581-6063 (online), ISSN:0972-5210

EFFECT OF USE OF THE AZOLLA PLANT INSTEAD PARTIAL FOR

SOYBEAN MEAL IN THE RATION OF COMMON CARP FISH CYPRINUS
CARPIO L. IN SOME PRODUCTION STANDARDS

Ammar A. Abd-Alnafi Al-Aaraji* and Nidhal T. Taha

Department of Animal Production, College of Agriculture and Forestry, Mosul University, Iraq.

Abstract
The soybean meal is an important source of protein in the feed ration of the common carp fish Cyprinus carpio L. and
requires its import from outside the country which increases the cost of the dietary ration of fish. The aim of this study is to
use Azolla sp. As a source of protein it is a partial substitute for soybean meal to reduce the cost of feeding the fish. The
experiment included feeding carp fish using five experimental diets containing Azolla in percentages (0, 2.5%, 5%, 7.5%, 10%)
of the total ration and Azolla substitutions (0, 15%, 30%, 45%, 60%) an alternative to soybean meal, and the use of glass
ponds in a 56-day carp growth experiment. The results of the statistical analysis of the weight gain values (g/fish) showed
that there were significant differences (P0.05) for the experimental treatments and the second treatment (Azolla 2.5%) was
high significance compared to the rest of the treatments, while the values of the growth rate and the relative growth rate% and
specific growth rate to There were no significant differences in the treatments (1, 2, 3, 4) respectively and differed significantly
with the fifth treatment. The results of the statistical analysis in the values of the feed conversion ratio and the percentage of
feed efficiency ratio% indicated that there were no significant differences between the second treatment (Azolla 2.5%) and
the first treatment (control) and the presence of significant differences (P0.05) with the rest of the treatments. The results of
the statistical analysis of the precipitated protein values, the protein efficiency ratio and the protein productive value%
confirmed that there were no significant differences for the treatments (1, 2, 3) respectively and differed significantly (P0.05)
with the fourth and fifth treatment. It is inferred that the Azolla can be substituted as a partial substitute for soybean meal
(15%) as the best growth criterion.
Key words: Azolla, Soybean meal, Ration, Cyprinus carpio L.

Introduction enter in many countries of the world (Khan et al., 2016),

its ability to accept and use grains in its nutrition and
Fish is an inexpensive food for the consumer with a
rapid daily growth reaches up to (2-4%) of body weight,
high nutritional value because it is rich in proteins,
The fish weighs (0.6 - 1.0) kg during one season in the
unsaturated fatty acids, phosphorus and calcium
ponds. Feeding fish accounts for more than 50% of the
(Dallinger et al., 1987) and after increasing health
cost of cultivation and requires finding substitutes for
awareness of the value of fish in nutrition and increasing
expensive feed materials such as soybean meal with plant
population and overfishing of fish and marine pollution
sources of protein that are inexpensive. One of these
led to increased demand for fish for the purpose of dam
alternatives is Azolla sp. (Sajid et al., 2016).
The local market needs of fish and the search for
alternatives to fish production. Hence the development Azolla contains about 19-30% raw protein on a dry
of fish farming started by raising fish especially the weight basis when conditions are appropriate for growth
common carp Cyprinus carpio L. which is the most (Peters et al., 1979 and Becking, 1979), and also contains
common fish species living in freshwater bodies lakes vitamin A, vitamin B12 and beta-carotene which stimulate
dams and streams (Vilizzi et al., 2015). The growth of a fish growth (Pillai et al., 2002) The research involved
good carp fish and the ability to withstand the harsh replacing Azolla as an alternative to soybean meal.
environment and its adaptation to industrial feeds made it The study aimed to find out the effect of using the
*Author for correspondence : E-mail : [email protected] Azolla plant in different proportions partly as a substitute
5178 Ammar A. Abd-Alnafi Al-Aaraji and Nidhal T. Taha

for soybean meal in experimental carp ration on some

production criteria. EXTECH D0600 field device at a rate of 5.5 mg/l, and
the pH level was 7-7.3 using the LABTECH (DIGITAL
Materials and Methods pH METER) device. It is within the recommended limits
The study was conducted in the fish laboratory of (FAO, 1981).
the College of Agriculture and Forestry, University of The five experimental rations in the fish laboratory
Mosul for a period of ten weeks including a two-week were made from feed materials that were ground by a
acclimatization period using glass basins of dimensions German origin laboratory mill, by replacing the Azolla
(40×60×40 cm) and equipped each tank with a Chinese- plant in part as a substitute for soybean meal in different
type (RS-510) air pump, with feeding all AUTOSAN type proportions (0%, 2.5%, 5%, 7.5%, 10%) for treatments
air compressor tubs of Chinese origin. (1, 2 , 3, 4, 5) respectively, shown in table 1. The
150 fish were distributed at an average weight of proportions of the feed materials were mixed well for
8.72 ± 0.05 g/fish in 15 glass basins with ten fishes/ponds, the purpose of homogeneity of the mixture for each ration
and with three replicates/treatment, the fish remained in separately and a cup of warm water was added to the
the ponds for two weeks to adapt them to the laboratory mixture and then the mixture was placed in the national
and pond environment and learn to eat food. meat grinding machine (Japanese origin with 4 mm holes)
gave small and coherent prints and dried for three days
These basins were equipped with liquid water from
cut into small pieces Fits the size of the mouth of the
a large water tank inside the laboratory in which the water
experimental fish and placed in opaque bags to prevent
was stored for 24 hours to ensure that it is free of chlorine
exposure to light and kept in plastic boxes. The
and obtain water at a moderate temperature. The
replacement process was performed as follows:
laboratory temperature was controlled between (25-30)
Celsius using air conditioners. 1- Control ration (1): free from Azolla.
It disposes of waste excreted and food residues in 2- Ration (2): It contains Azolla 4.5% and soybean meal
ponds on a daily basis by partial substitution of siphon 27.5% of total ration, that is a substitution rate 15% of
water by 20-25% and supplementing water from the tank, the soybean meal.
the fish are fed twice daily during the adaptation period 3- Ration (3): It contains Azolla 9% and soybean meal
until the start of the research experiment. 25% of total ration, that is a substitution rate 30% of
The water temperature of the glass ponds was the soybean meal.
measured by a mercury thermometer and was at a 4- Ration (4): It contains Azolla 13.5% and soybean meal
temperature of 25°C It was the appropriate temperature 22.5% of total ration, that is a substitution rate 45% of
for common carp growth (Crockford and Johnston, 1990). the soybean meal.
Dissolved oxygen was measured in ponds using an 5- Ration (5): It contains Azolla 18% and soybean meal
Table 1: Composition of experimental rations components (%) resulting 20% of total ration, that is a substitution rate
from substituting different proportions of Azolla as a partial 60% of the soybean meal.
substitute for soybean meal.
Experiment fish were fed on experimental
Experimental (1) (2) (3) (4) (5) rations mentioned in table 1 at a rate of 3-5%
rations Azolla Azolla by Azolla by Azolla by Azolla by of body weight, two-meal ration per day was
Ingredients (zero) substitu- substitu- substitu- substitu- adopted and the amount of ration provided to
used control tion 15% tion 30% tion 45% tion 60% experiment fish was increased based on the
Fish meal 10 10 10 10 10 weight gain of fish growth, with fish weights
Soybean meal 30 27.5 25 22.5 20 taken every two weeks using electronic
Azollam --- 4.5 9 13.5 18 sensitive balance (0.01)g Chinese type citizen
Barley 20 20 20 20 20 for eight weeks, and the feed provided to the
Yellow corn 18.5 16.5 14.5 12.5 10.5 fish was cut one day per week to increase the
Bran 19 19 19 19 19 fish’s appetite to eat the feed for the next day
Salt 0.5 0.5 0.5 0.5 0.5 during the trial period.
Vitamins 1 1 1 1 1
Methods for measuring the growth
Limestone 0.5 0.5 0.5 0.5 0.5
Bonding article 0.5 0.5 0.5 0.5 0.5
criteria for experiment fish:
Total 100 100 100 100 100 The criteria for measuring fish growth
 Use of the azolla plant instead partial for soybean meal in the ration of common carp fish cyprinus carpio L. 5179

calculations were adopted to show the effect of Azolla

plant substitutes in part on soybean meal in their growth Feed consumption ( g )
represented by the total weight gain of fish (T.W.G) Feed conversion ratio =
(Pitcher and Hart, 1982) and the growth rate of fish Total weight gain ( g )
Growth Rate (G.R) (Pitcher and Hart, 1982), Relative Feed efficiency ratio (%) =
Growth Rate (R.G.R)% (Uten, 1978), Specific Growth
Rate (S.G.R) (Jobling and Koskela, 1996) and Feed Total weight gain ( g )
Conversion Ratio (F.C.R) (Uten, 1978), Feed Efficiency Feed consumption ( g ) × 100
Ratio (F.E.R) (Uten, 1978), Protein Efficiency Ratio
(P.E.R) (Gerking, 1971), Protein Intake and Protein Total weight gain ( g )
Productive Value (P.P.V) (Jobling and Koskela, 1996), Protein efficiency ratio =
Pr otein int ake ( g )
according to the equations:
Total weight gain (g/fish) = Final weight (g) – Initial Protein productive value (%) =
weight (g)
Pr otein gain ( g )
Daily weight gain (g/fish) =
Pr otein int ake ( g ) ×100
Final weight  Initial weight Protein gain (g) = Body protein final of experiment
Number of days (g) - Body protein initial of experiment (g)m
Daily growth rate (g/fish/day) = Precipitated protein = % The crude protein in the
fish’s body for final weight - % The crude protein in the
Weight gain ( g ) fish’s body for the initial weight
Duration of exp eriment ( days) Chemical analyzes of fish growth experiment:
(components of nutrients):
Relative growth rate (%) =
Determination of nutrient components (protein, fat,
Final weight ( g )  Initial weight ( g ) ash, moisture, and nitrogen free extract) in the dry matter
× 100 was performed:
Initial weight ( g / fish )
1- Ingredients of experimental processed fish rations
Specific growth rate = shown in table 2.
Log final weight ( g )  Log initial weight ( g ) 2- The eaten part of the fish of the experiment
Duration of exp eriment ( days ) research.

Table 2: The chemical composition (%) of the five experimental rations The nutrients in fish rations and body were
calculated on the basis of the dry weight in which the protein estimated on a dry weight basis according to
ratio was established between (26.38-28.13) for the purpose of the standard methods approved (AOAC, 2000)
showing the effect of Azolla on the growth of experimental fish. in estimating the protein, fat, ash and moisture,
Experimental (1) (2) (3) (4) (5) according to the nitrogen free extract
rations Azolla Azolla by Azolla by Azolla by Azolla by mathematically using the difference method as
Ingredients (zero) substitu- substitu- substitu- substitu- shown in (Wee and Shu, 1989) as follows:
used control tion 15% tion 30% tion 45% tion 60% Dissolved carbohydrates = 100 - (crude
Humidity 7.75 6.56 7.05 6.55 6.91 protein% + fat% + ash% + fiber% +
Ash 5.68 6.39 6.94 7.77 7.99 moisture%).
Raw fat 6.26 7.53 6.69 6.39 6.92 Statistical analysis
Raw protein 26.38 25.68 26.38 27.13 28.13
Nitrogen free 53.93 53.84 52.94 52.16 50.05
Data analysis was performed statistically
extract(NFE) using complete randomized design (CRD) by
Total 100 100 100 100 100 statistical package for social science (SPSS,
*Metabolicm 14.49 14.78 14.51 14.44 14.51 2001) in analyzing the effect of experimental
energy (MJ/kg) treatments and testing significant differences
between mean characteristics attained by
*Metabolic energy was calculated based on Smith, (1971) equation: ME(MJ/
Duncan´s multiple rang test (Duncan, 1955).
Kg) = Protein × 18.8 + Fat × 33.5 + NFE × 13.8.
5180 Ammar A. Abd-Alnafi Al-Aaraji and Nidhal T. Taha

Results and Discussion Sahu, (2006) reported that Azolla plant is a good
Criteria for growth, weight gain and growth rate source of protein and can be used up to 16.25% by
substituting 10% as a substitute for fish meal in the feed
The results of the statistical analysis of the final weight rations of Labeo rohita Fry fish, and Azolla substitution
rate (g/fish) in table 3 showed that there were no can reach 25% for rohita fish feeding (Datta, 2011).
significant differences (P0.05) between the treatments
(1, 2, 3, 4) respectively, while a significant difference It was observed in our current research that an
was observed for the fifth treatment (Azolla 10%) it increase in the percentage of Azolla substitution of 15%
reached (15.07) g/fish. The results of the statistical in the experimental rations of common carp leads to a
analysis of the weight gain (g/fish) showed that there decrease in the weight gain rate and growth rate, and
were significant differences (P0.05) for the experimental this is what Sahu pointed out, (2006), that the decrease in
treatments and the second treatment (Azolla 2.5%) growth due to the increased concentration of Azolla in
whose value was (8.44) g/fish with high significance dietary rations is due to unbalance of amino acid
compared to the rest of the treatments, where they composition in Azolla protein, Azolla values were higher
differed significantly with The third treatment (Azolla with methionine and lysine amino acids compared to
5%) and its value was (7.85) g/fish, and the difference soybean meal and meat and bone powder (Das et al.,
was highly significant with the fourth treatment (Azolla 2018). Our research agreed with Gangadhar et al., (2015)
7.5%) and the fifth treatment (Azolla 10%) which when adding different levels of Azolla in Labeo
amounted to (7.77 , 6.20) g/fish respectively, while No fimbriatus fish, and researcher Cruz-Velásquez et al.,
significant differences were observed between the third (2014) showed similar results to our research, as well as
treatment (Azolla 5%) and control. The results of the the compatibility of results with Micha et al., (1988) where
growth rate (g/fish/day) in table 3 showed that there were he mentioned a decrease Fish (Oreochromis niloticus
no significant differences for the four treatments (1, 2, 3, and Tilapia rendalli) grow when adding Azolla to feeding
4) compared to the fifth treatment (Azolla 10%) which rations. The differences in the criteria for fish growth
amounted to (0.11) g/fish/day. are also attributed to the different ratios of Azolla added
to the ration that vary the energy ratios of
Table 3: Effect of different levels of Azolla sp. on the growth criteria,
experimental feeds (Lupatsch, 2001).
weight gain and growth rate of common carp fed for 56 days
(mean ± standard error). Relative and specific growth rate
Studied Initial Final Weight Growth The results of the statistical analysis in table
criteria weight weight gain rate 4 showed that the relative growth values showed
Treatments (g/fish) (g/fish) (g/fish) that there were no significant differences for
(g/fish)
Control (1) 8.46 ± 0.12a 16.68 ± 0.19a 8.22 ± 0.08ab the treatments (1, 2, 3, 4) respectively while a
0.15 ± 0.01a
Azolla (2.5%) (2) 8.69 ± 0.40a 17.14 ± 0.19a 8.44 ± 0.219a significant difference was observed in the fifth
0.15 ± 0.01a
Azolla (5%) (3) 9.18 ± 0.28a 17.03 ± 0.31a 7.85 ± 0.32ab treatment (Azolla 10%) compared to the rest
0.14 ± 0.01a
Azolla (7.5%) (4) 8.92 ± 0.27a 16.69 ± 0.27a 7.77 ± 0.10b of the treatments which amounted to (69.95%).
0.14 ± 0.01a
Azolla (10%) (5) 8.87 ± 0.07a 15.07 ± 0.01a 6.20 ± 0.08c The results of the statistical analysis of the
0.11 ± 0.01b
*The various letters within the single column of the studied trait indicate specific growth values indicated that there were
significant differences at probability level (P0.05). no significant differences for the treatments (1,
2, 3, 4) respectively while a significant difference was
Table 4: Effect of different levels of Azolla sp. on the criteria
for relative growth and specific growth of common
observed in the fifth treatment (Azolla 10%) compared
carp fed for 56 days (mean ± standard error). to the rest of the treatments and its value (1.16).

Studied criteria Specific Relative

It was observed during the current research that the
Treatments growth growth % relative growth rate% and specific growth in feeding
Control (1) 97.22 ± 0.79a 1.21 ± 0.01a rations using 15% Azolla and Azolla free control rations
Azolla (2.5%) (2) 97.72 ± 6.57a 1.22 ± 0.01a were more appreciated than other treatments, and this
Azolla (5%) (3) 85.84 ± 5.12a 1.21 ± 0.01a was agreed with Micha et al., (1988, 1989) and El-Sayed,
Azolla (7.5%) (4) 87.29 ± 3.13a 1.21 ± 0.01a (1992), The researchers observed that when adding
Azolla (10%) (5) 69.95 ± 1.41b 1.16 ± 0.01b Azolla to the fish O. niloticus feed it reduced the growth
rate. This decrease in growth is linked to a decrease in
*The various letters within the single column of the studied
the intake of food by regular carp and trout (Ogino, 1980),
trait indicate significant differences at probability
level (P0.05). and the lack of nutrition reduces the rate of fish growth,
 Use of the azolla plant instead partial for soybean meal in the ration of common carp fish cyprinus carpio L. 5181

Increased Azolla reduced the digestion and growth rate the feed conversion ratio indicated that there were no
in the ration due to the difficulty of digesting raw vegetable significant differences in the first and second treatment
ingredients Because the Azolla plant contains large (Azolla 2.5%) and their values (2.58 , 2.57) respectively,
quantities of non digestible fibers and is generally with a highly significant difference (P0.05) for the fifth
considered to be complex molecules represented by treatment (Azolla 10%) reached Its value (3.38) and this
cellulose and starch (De Silva et al., 1995), because value is high compared to the third treatment (Azolla
cellulose is difficult to digest by fish while starch is 5%) and the fourth (Azolla 7.5%) which differed morally,
converted to glucose and in turn saves energy. Fagbenro and we did not notice a significant difference between
et al., (2004) and Omoregie et al., (1993) explain that the third transactions (Azolla 5%) and the fourth (Azolla
increased fiber content in dietary rations of fish and 7.5%) whose value was (2.76, 2.91) respectively. The
vegetable sources has a negative effect on weight gain, results of the statistical analysis in the values of feed
protein representation and growth response in Nile tilapia, efficiency ratio% showed that there were no significant
Anderson et al., (1984) noted when the fiber level differences in the first treatment and the second treatment
exceeds 100 g/kg reduced the feeding efficiency and (Azolla 2.5%) and differed significantly from the third
digestion of tilapia fish resulting in poor fish growth. As treatment (Azolla 5%) and the fourth (Azolla 7.5%),
well as the Azolla containing anti-nutritional factors that while no significant differences were recorded between
may adversely affect the efficiency of fish feeding the third treatment (Azolla 5%) and the fourth (Azolla
(Krogdahl et al., 2010). 7.5%), while a highly significant difference (P0.05) was
Fasakin, (1999) indicated that the Azolla africana observed between the first and the second treatment
plant contains a small fraction of some anti-nutritional (Azolla 2.5%) compared to the fifth treatment (Azolla
factors such as cyanide, tannin and phytic acid. Micha et 10%) and differed significantly with the third and fourth
al., (1998) observed that the growth rate of tilapia fish treatment.
decreased gradually with the increase in the level of the The results of the feed conversion ratio announced
Azolla microphylla plant it is evident from the current by Fiogbé et al., (2004) indicated that the observed
research that the growth performance of fish is due to growth difference was due to insufficient levels of amino
the lack of quality protein, amino acids, fats and energy acids and caused a reduction in food intake, protein
digestion of the plant the nutritional quality of the Azolla synthesis and growth. This was confirmed by Cowey
plant can be improved by adding chemical and mechanical (1992), Cole and van Lunen (1994).
treatments it concentrates the protein content and removes The researchers observed Almazan et al., (1986)
the effects of anti-nutritional substances but it will increase with O. niloticus, Antoine et al., (1986) with O.
the cost of feed. niloticus, Cichlasoma melanurum and Micha et al.,
Food intake, feed conversion ratio and feed (1988) with O. niloticus and T. rendalli and Joseph et
efficiency ratio al., (1994) with Etroplus suratensis fish Decreased
The results of the statistical analysis in table 5 of the growth performance and feed conversion ratio when
amount of food intake (g/fish) showed that there were increasing Azolla supplementation in nutrition. A similar
no significant differences between the different decrease in growth rate and feed conversion ratio was
treatments. While the results of the statistical analysis of observed in fish feeding on high levels of Azolla
concentration and it was found that the lower growth
Table 5: Effect of different levels of Azolla sp. on the food intake was due to an imbalance in the composition of the
criteria, feed conversion ratio and feed efficiency ratio of amino acid of the Azolla protein (Sahu and Sheeno,
common carp fed for 56 days (mean ± standard error).
2006).
Studied criteria Feed efficie- Feed conv- Food intake Almazan et al., (1986) indicated that the Azolla
Treatments ncy ratio% ersionratio (g/fish) pinnata protein is limited to a small amount of
Control (1) 21.16 ± 1.15a 2.58 ± 0.15b 39.10 ± 2.19a tryptophan amino acid and a severe deficiency of
Azolla (2.5%) (2) 21.62 ± 1.18a 2.57 ± 0.21b 39.37 ± 3.02a threonine amino acid, causing confusion in increasing
Azolla (5%) (3) 21.58 ± 0.60a 2.76 ± 0.17ab 36.51 ± 2.33ab feed consumption for the purpose of compensating
Azolla (7.5%) (4) 22.60 ± 0.65a 2.91 ± 0.10ab 34.45 ± 1.25ab
for the deficiency of amino acids when increasing the
Azolla (10%) (5) 20.91 ± 1.46a 3.38 ± 0.28a 29.98 ± 2.33b
Azolla concentration in feed rations.
*The different letters within the single column of the studied trait
The reason for the increase in the feed conversion
indicate significant differences at the probability level
(P0.05). ratio when increasing the concentration of Azolla
5182 Ammar A. Abd-Alnafi Al-Aaraji and Nidhal T. Taha

compared to the feed rations free from Azolla is due to (Azolla 10%), no significant difference was observed
the presence of high dietary fiber and ash content, where between the fourth treatment (Azolla 7.5%) and the fifth
the highest percentage of the feed conversion ratio of (Azolla 10%). The highest protein productive value for
Nile tilapia fish (4.2) which was fed on food rations the second treatment (25.78%) and the lowest protein
fortified with dried Azolla (A. pinnata) at a level of 25% productive value was for the fifth treatment (19.67%).
(Abdel-Tawwab, 2008), a decline in feed digestion and The significant decrease in protein productive value
consumption occurs at the inclusion of high levels of Azolla %, precipitated protein and protein efficiency ratio
by affecting the activity of digestive system enzymes and indicates an increase in the level of Azolla our current
thus growth performance is linked to improved nutrient research agreed with Micha et al., (1988) and El-Sayed,
digestion and nutritional efficiency (Hong et al., 2004, (1992). (El-Sayed, 1992) explained that the lowest
Sithara et al., 2008). percentage can be replaced by Azolla 25% as a substitute
for fish meal in the ration, because adding Azolla to fish
Protein intake, protein efficiency ratio, precipitated feed rations depends on the cost / benefit ratio, taking
protein and protein productive value into account the differences between the types of fish,
The results of the statistical analysis in table 6 for their ages and their ability to digest Plant material and
the values of the protein intake (g/fish) showed no the nature of fish feed whether plant or animal or mixed
significant differences between the different treatments. (Buddington, 1979 and Caulton, 1978).
While the results of the statistical analysis of the protein Increasing the concentration of Azolla with a
efficiency ratio values indicated that there were no decrease associated with the soybean meal leads to a
significant differences for the treatments (1, 2, 3) lack of essential amino acids and consequently a decrease
respectively while a significant difference (P0.05) was in protein quality, as Santiago et al., (1988) and Olvera-
observed for the treatments (1, 2, 3) compared to the Novoa et al., (1990) indicate that this phenomenon has
fifth treatment (Azolla 10%) they differed significantly found similar results in the use of alfalfa and Azolla
with the fourth treatment (Azolla 7.5%) which amounted pinnata in nutritional ration for tilapia. Fasakin and
to (1.27). The results of the statistical analysis of the Balogun (1998) explained when precipitated protein and
precipitated protein values confirmed that there were no protein efficiency ratio decreased as the level of Azolla
significant differences for the first, second and third in the feeding ration of Clarias gariepinus increased.
treatment respectively while a significant difference with Our current research agreed with Hossain et al., (2002)
the fourth treatment (Azolla 7.5%) and highly significant and Fasakin et al., (2001) replacing fish meal with
difference (P0.05) with the fifth treatment (Azolla 10%) vegetable protein in the dietary rations of Nile tilapia.
appeared and the highest value for the precipitated protein
The high fiber content speeds food through the
was for the second treatment (Azolla 2.5%) and reached
(1.42%), while the lowest value for the precipitated protein intestines of animals (Bender, 1967) which reduces the
time available to digest and absorb nutrients. There is a
is for the fifth treatment (Azolla 10%) and it reached
(1.15%). The results of the statistical analysis of the negative relationship between the activity of the protease
enzyme and the concentration of the Azolla protein for
criteria for the protein productive value % showed that
there were no significant differences for the first three feed, it is mentioned when changes occur in the activity
of protein hydrolysis enzymes in common carp fish that
treatments respectively and differed significantly with the
fourth treatment (Azolla 7.5%) and the fifth treatment depend on the composition of dietary rations, as the
Azolla-free food may be more adapted to the
Table 6: Effect of different levels of Azolla sp. on the criteria of the protein protein hydrolysis enzyme which gradually
intake, protein efficiency ratio, precipitated protein, and protein deteriorates due to increased Azolla. Protein
productive value of common carp fed for 56 days (mean ± standard degradation activity may have decreased due
error).
to trypsin inhibitors present in the vegetable
Studied Protein Protein Precipi- Protein tissues of many aquatic plants (Gleen et al.,
criteria intake efficiency tated productive 1982 and Yousif et al., 1994) as trypsin inhibitors
Treatments (g/fish) ratio protein value % impair protein digestion and absorption (Olvera-
Control (1) 5.58 ± 0.30a 1.48 ± 0.08a 1.40 ± 0.01a 25.26 ± 1.16a Novoa et al., 1990).
Azolla (2.5%) (2) 5.55 ± 0.30a 1.53 ± 0.12a 1.42 ± 0.04a 25.78 ± 1.40a Chemical composition of fish body
Azolla (5%) (3) 5.69 ± 0.16a 1.39 ± 0.09a 1.40 ± 0.02a 24.68 ± 0.51a
The results of the statistical analysis of the
Azolla (7.5%) (4) 6.13 ± 0.18a 1.27 ± 0.05ab 1.30 ± 0.01b 21.30 ± 0.66b chemical composition of the fish body table 7
Azolla (10%) (5) 5.88 ± 0.41a 1.07 ± 0.08b 1.15 ± 0.01c 19.67 ± 1.20b of the values of humidity percentage for the
*The various letters within the single column of the studied trait indicate components of the eaten part of the fish
significant differences at probability level (P0.05). indicated that there were significant differences
 Use of the azolla plant instead partial for soybean meal in the ration of common carp fish cyprinus carpio L. 5183

Table 7: Effect of different levels of Azolla sp. on the chemical composition of the common carp fish body (mean ± standard
error).
Treatments The chemical composition of fish body% after feeding it on different experimental rations
Studied Before Control Azolla Azolla Azolla Azolla
criteria the (zero) (2.5%) (5%) (7.5%) (10%)
experiment (1) (2) (3) (4) (5)
Humidity abc72.17± 0.13 bc72.07± 0.35 c72.01± 0.05 a72.50± 0.16 ab72.43± 0.12 abc72.28± 0.01
Dry matter abc27.83± 0.13 ab27.93± 0.11 a27.99± 0.05 c27.50± 0.16 bc27.57± 0.12 abc27.72± 0.10
Crude protein c15.32± 0.09 b16.23± 0.07 b16.27± 0.04 ab16.47± 0.12 ab16.53± 0.12 a16.63± 0.16
Raw fat a7.11±0.08 ab6.81±0.15 abc6.74±0.05 bc6.64±0.24 cd6.38±0.08 d6.22±0.05
Ash ab3.82±0.17 b3.71±0.13 ab3.86±0.01 c3.32±0.18 bc3.67±0.01 a4.12±0.04
Carbohydrates a1.58±0.05 b1.18±0.22 b1.12±0.03 bc1.07±0.02 bc0.99±0.07 c0.75±0.15
*The different letters within the same row of the studied trait indicate significant differences at the probability level (P0.05).

(P0.05) between the different treatments where the body. El-Sayed, (1992) explained that body protein and
percentage of humidity in the third treatment (Azolla 5%) fat were negatively associated with high levels of Azolla
reached (72.50) from the humidity content of the fish in fish rations while the association was positive for ash
body prior to the first trial and treatment (control) was and body humidity. Raw body protein data in our current
(72.17, 72.07) respectively. The results of the statistical study contrasted with the illustration he mentioned (El-
analysis of the percentage values of the dry matter Sayed, 1992) but by agreement regarding the fat, ash
percentage of the components of the eaten part of the and body humidity content (Micha et al., 1988) that the
fish showed that there were significant differences addition of Azolla at increased levels in fish rations of O.
(P0.05) between the different treatments and the highest niloticus and T. rendalli resulted in a reduction in the
value of the second treatment (Azolla 2.5%) and reached percentage of fats in the body tissues of both species
(27.99). The results of the statistical analysis showed the and no effect on the crude protein content.
values of the crude protein ratio to the presence of highly Das, (2018) stated when adding different levels of
significant (P0.05) for the fifth treatment (Azolla 10%) Azolla to the Thai silver Barbonymus gonionotus feed
and its value (16.63) for the ratio of crude protein before ration that raw fat, protein and ash were negatively
the experiment and reached (15.32) and the first treatment correlated while the correlation was positive relative to
amounted to (16.23), while it was not recorded significant humidity.
differences between the second treatment (Azolla 2.5%)
and the first treatment. Likewise, no significant difference Conclusion
was recorded between the fourth treatment (Azolla 1- Azolla is a good source of protein and can be replaced
7.5%) and the third treatment (Azolla 5%). The results in rations for common carp fish by 15% partly as a
of the statistical analysis of the raw fat percentage values substitute for soybean meal without any harmful
indicated the presence of significant differences (P0.05) effect.
among the different treatments, significant differences
were observed (P0.05) for the ash percentage values 2- Adding Azolla to fish rations reduced the percentage
between the different treatments and the highest ash ratio of fat in fish muscles.
value was in the fifth treatment (Azolla 10%) and its 3- Azolla is an inexpensive protein source compared to
value was (4.12) and the lowest value was for the third other protein sources that require import.
treatment (Azolla 5%) and reached (3.32). From the table 4- Azolla plant productivity is characterized by its high
7 in which the results of the statistical analysis of the
rate of growth and its tolerance to harsh environmental
carbohydrate percentage values were found there were
conditions.
significant differences (P0.05) between the different
treatments and no significant differences were observed Acknowledgments
between the first treatment (control) and the second
I would like to express my deepest gratitude and
treatment (Azolla 2.5%) as well as the absence of
significant differences between the third treatment (Azolla supervisor ‘’Dr. Nidhal Tahseen Taha’’ for his helper
5%) and the fourth treatment (Azolla 7.5%). advices, meticulous supervision, and guidance throughout
the execution of this research.
Antoine et al., (1986) indicated when feeding O.
niloticus and C. melanurum fish on Azolla they had high My special thanks and gratitude to my parent and
humidity content and low fat concentrations in the fish their continued support in life, my beloved wife, all
5184 Ammar A. Abd-Alnafi Al-Aaraji and Nidhal T. Taha

members of my (University of Mosul, Collage of Datta, S.N. (2011). Culture of Azolla and its efficacy in diet of
Agriculture and Forestry) and all my friends. Labeo rohita. Aquaculture, 310(3-4): 376-379.
De Silva, S.S. and T.A. Anderson (1995). Fish Nutrition in
References Aquaculture. Chapman & Hall Aquaculture, London, UK,
A.O.A.C. Association of official analytical chemists (2000). 17 319 pp.
th
ed. V11. USA. Duncan, C.B. (1955). Multiple rang and Multiple “ F “ test.
Abdel-Tawwab M. (2008). The preference of the omnivorous- Biometric, 11: 1-12.
macrophagous. Tilapia zillii. El-Sayed, A.F.M. (1992). Effect of substituting fish meal with
Gervais. to consume a natural free-floating fern. Azolla pinnata. Azolla pinnata in practical diets for fingerling and adult
Journal of the World Aquaculture Society, 39(1): 104- Nile tilapia, Oreochromis niloticus L. Aquacult. Fish.
112. Manag, 23: 167-173.
Almazan, G.J., R.S.V. Pullin, A.F. Angels, T.A. Manolo, R.A. Fagbenro, O.A., T.T. Akande, T.T. Fapohunda and Y. Akegbejo-
Agbayani and M.T.B. Trono (1986). Azolla pinnata as Samsons (2004). Comparative assessment of roselle
dietary component for Nile tilapia, Oreochromis niloticus. Hibiscus sabdariffa var. sabdariffa seed meal and kenaf
In J.L. Maclean, L.B. Dizon and L.V. Hosillos Eds., The Hibiscus sabdariffa var. altissima seed meal as replacement
first Asian fisheries forum proceedings pp: 523-528. Manila: for soybean meal in practical diets for fingerlings of Nile
Asian Fisheries Society. Tilapia. Oreochromis niloticus. Pages 277-288 in R. Bolivar,
G. Mair and K. Fitzsimmons. editors. The 6th International
Anderson, J., A.J. Jackson, A.J. Matty and B.S. Capper (1984). Symposium of Tilapia in Aquaculture. Manila. The
Effects of dietary carbohydrate and fiber on the Philippines; September 14-16. 2004.
Tilapia Oreochromis niloticus L. Aquaculture, 37: 303-
314. Fasakin, A.E. and A.M. Balogun (1998). Evaluation of the dried
water fern Azolla pinnata as a replacer of soybean dietary
Antoine, T., S. Carraro, J.C. Micha and C. Van Hove (1986). components for Clarias gariepinus fingerlings.  J.
Comparative appetency for Azolla of Cichlasoma and Aquacul. Trop., 13: 57-64.
Oreochromis tilapia. Aquaculture, 53: 95-99.
Fasakin, E.A. (1999). Nutrient quality of leaf protein
Becking, J.H. (1979). Environmental requirements of Azolla for concentrates produced from water fern Azolla africana
use in tropical rice production. In: Nitrogen and Rice. Desv and duckweed Spirodela polyrrhiza L. Schleiden.
International Rice Research Institute, Los Banos, Laguna, Biores. Technol., 69: 185-187.
Philippines. pp: 345-374.
Fasakin, E.A., A.M. Balogun and O.A. Fagbenro (2001).
Bender, A.E. (1967). Dietetic Foods. Leonard Hill Books, Evaluation of Sun-Dried Water Fern, Azolla Africana and
London, pp: 211. Duckweed, Spirodela polyrrhiza in Practical Diets for Nile
Buddington, R.K. (1979). Digestion of an aquatic macrophyte tilapia, Oreochromis niloticus Fingerlings. J. Appl.
by Tilapia zillii Gervais. J. Fish. Biol., 15: 449-455. Aquac., 11: 83-92.
Caulton, M.S. (1978). Tissue depletion and energy utilization Fiogbé, E.D., J.C. Micha and C. Van Hove (2004). Use of a
during routine metabolism by sub-adult Tilapia rendalli natural aquatic fern, Azolla microphylla, as a main
Boulenger. J. Fish. Biol., 13: 1-6. component in food for the omnivorous–phytoplankton
Cole, D.J.A. and T.A. Van Lunen (1994). Ideal amino acid Ophagous tilapia, Oreochromis niloticus L. Journal of
patterns. In: Amino Acids in Farm Animal Nutrition. J.P.I. Applied Ichthyology, 20(6): 517-520.
D’Mello (Ed.). The Scottish Agricultural College, Food and Agriculture Organization (FAO) (1981). Report of the
Edinburgh, UK, pp: 99-112. symposium on new developments in the utilization of the
Cowey, C.B. (1992). Nutrition: estimation requirements of heated effluents in the circulation system for intensive
rainbow trout. Aquaculture, 100: 177-189. aquaculture stavanger, 29-30, Rome. Italy.
Crockford, T. and I.A. Johnston (1990). Temperature acclimation Gangadhar, B., N. Sridhar, S. Saurabh, C.H. Raghavendra, K.P.
and the expression of contractile protein isoforms in the Hemaprasanth and P. Jayasankar (2015). Effect of Azolla
skeletal muscles of the common carp Cyprinus carpio L. incorporated diets on the growth and survival of Labeo
Journal of comparative physiology B, 160(1): 23-30. fimbriatus during fry-to fingerling rearing. Cogent Food
and Agriculture, 1: 1055539.
Cruz-Velásquez, Y., C. Kijora, W. Vergara-Hernández and C.
Schulz (2014). On-farm evaluation of Cachama blanca Gerking, S.D. (1971). Influence of rate of feeding and body
and Nile tilapia fed fermented aquatic plants in a weight on protein metabolism of bluegill sunfish.
polyculture. Orinoquia Suplemento, 18(2): 269-277. Physiological Zoology, 44(1): 9-19.þ
Dallinger, R., F. Prosi, H. Segner and H. Back (1987). Gleen, E.P., M.R. Fontes, M.R. Katzen and L.B. Colvin
Contaminated food and uptake of heavy metals by fish: a (1982). Nutritional Value of Halophytes Grown on Hyper
review and a proposal for further research. Oecologia, Saline Sea Water. In: Biosaline Research: A Look to the
73(1): 91-98. Future, Sanpietro, A. (Ed.). Plenum Press, New York, pp:
485-489.
Das, M., Rahim, F.I. & Hossain, A.,.2018. Evaluation of Fresh
Azolla pinnata as a Low-Cost Supplemental Feed for Thai Hong, K.J., C.H. Lee and S.W. Kim (2004). Aspergillus oryzae
Silver Barb Barbonymus gonionotus. Fishes, 3(15): 1-11. GB-107 fermentation improves nutritional quality of food
doi:10.3390/ûshes3010015. soybeans and feed soybean meals. J. Med. Food, 7: 430-
 Use of the azolla plant instead partial for soybean meal in the ration of common carp fish cyprinus carpio L. 5185

435. as a protein source in diets for tilapia Oreochromis

Hossain, M.A., U. Focken and K. Becker (2002). Nutritional mossambicus. Aquaculture, 90: 291-302.
evaluation of dhaincha (Sesbania aculeata) seeds as Omoregie E. and F.I. Ogbemudia (1993). Effect of substituting
dietary protein source for tilapia Oreochromis niloticus. fishmeal with palm kernel meal on growth and food
Aquac. Res., 33: 653-662. utilization of the Nile Tilapia. Oreochromis niloticus.
Jobling, M. and R. Koskela (1996). Inter-individual variation in Israeli Journal of Aquaculture – Bamidgeh, 45: 113-119.
feeding and growth in rainbow trout Oncorhynchus mykiss Peters, G.A., B.C. Mayne, T.B. Ray and R.E. Jr. Toia (1979).
during restricted feeding and in a subsequent period of Physiology and biochemistry of the Azolla-
compensatory growth. J. Fish. Biol., 49: 658-667. Anabaena symbiosis. In: Nitrogen and Rice. International
Joseph, A., P.M. Sherief and T. James (1994). Effect of different Rice Research Institute, Los Banos, Laguna, Philippines.
dietary inclusion levels of Azolla pinnata on the growth, pp: 325-344.
food conversion and muscle composition of Etroplus Pillai, P.K., S. Premalatha and S. Rajamony (2002). Azolla-A
suratensis Bloch. Journal of Aquaculture in the Tropics, sustainable feed substitute for livestock. Leisa India, 4(1):
9: 87-94. 15-17.
Khan, M.N., K. Shahzad, A. Chatta, M. Sohail, M. Piria and T. Pitcher, T.J. and P.J.B. Hart (1982). Fisheries Ecology. London
Treer (2016). A review of introduction of common carp and Candberra; Crom Helm Ltd. American edition: The Avi
Cyprinus carpio in Pakistan: Origin, purpose, impact and pubi Co., Linc. Westport, Conn. 414 pp.
management. Croat. J. Fish., 74: 71-80. Sahu, T.S.P. (2006). Use of freshwater aquatic plants as a
Krogdahl, A., M. Penn, J. Thorsen, S. Refstie and A.M. Bakke substitute of fishmeal in the diet of Labeo rohita fry.
(2010). Important antinutrients in plant feedstuffs for Journal of Fisheries and Aquatic Science, 1(2): 126-135.
aquaculture: An update on recent findings regarding Sajid, M., K. Noor, J.I. Khalid, A. Muhammad and K. Anjum
responses in salmonids. Aquac. Res., 41: 333-344. (2016). Evaluation of water hyacinth Eichhornia crassipes
l i ve s t o ck . h t tp : / / ww w. er i k s jo d i n .n e t / p r o j e ct s / supplemented diets on the growth, digestibility and
theAzollacookingandcultivationproject/ research/Azolla, histology of grass carp Ctenopharyngodon idella
a sustainable feed for livestock.pdf. fingerlings. J. Appl. Anim. Res., 46(1): 24-28.
Lupatsch, I., G.W. Kissil, D. Sklan and E. Pfeffer (2001). Effects Santiago, C.B., M.B. Aldaba, O.S. Reyes and M.A. Laron
of varying dietary protein and energy supply on growth, (1988). Response of Nile Tilapia Oreochromis niloticus
body composition and protein utilization in gilthead sea Fry to Diets Containing Azolla Meal. Proceedings of 2nd
bream Sparus aurata L. Aquaculture Nutrition, 7(2): 71- International Symposium on Tilapia Aquaculture, Mar. 16-
80. 20, Bangkok,Thailand, pp: 377-382.
Micha, J.C., T. Antoine, P. Wery and C. Van Hove (1988). Growth, Sheeno, T.P. and N.P. Sahu (2006). Use of freshwater aquatic
ingestion capacity, comparative appetency and plants as a substitute of fish meal in the diet of Labeo
biochemical composition of Oreochromis niloticus and rohita fry. Journal of Fisheries and Aquatic Science, 1:
Tilapia rendalli fed with Azolla. In R.S.V. Pullin, T. 126-135.
Bhukaswan, K. Tonguthai and J.L. Maclean (Eds.), Second
International Symposium on Tilapia in Aquaculture. Sithara, K. and K. Kamalaveni (2008). Formulation of low-cost
ICLARM Conference Proceedings pp: 347-355. Bangkok: feed using Azolla as a protein supplement and its influence
Department of Fisheries. on feed utilization in fishes. Current Biotica, 2: 212-219.
Micha, J.C., B. N’Guessan and C. Van Hove (1989). The aquatic Smith, R.G. (1971). A method for measuring digestibility and
fern Azolla as feed for fish. pp: 677-681. In: Aquaculture, metabolizable of energy of feeds. Prog. Fish Cult., 33:
A Biotechnology in Progress, 2: E. de Pauw, E. Jaspers, H. 132-134 .
Ackefors and N. Wilkins (Eds). European Aquaculture Statistical Package for Social Science (2001). SPSS Version 10,
Society, Bredene, Belgium. SPSS Inc, U.S.A.
Micha, J.C., T. Antoine, P. Wery, Van Hove and C. Growth Uten, F. (1978). Standard methods and terminology in fin fish
(1998). Ingestion capacity, comparative appetency and nutrition from: proc. World Sump. On Fin Fish nutrition
biochemical composition of Oreochromis niloticus and and fish feed technology. Hamburg, 20-23. June 1978. Vol.
Tilapia rendalli fed with Azolla. In Proceedings of the 11 Berlin.
Second International Symposium on Tilapia in Vilizzi, L., A.S. Tarkan and G.H. Copp (2015). Experimental
Aquaculture, Department of Fisheries, Bangkok, Thailand, evidence from causal criteria analysis for the effects of
16-20 March 1987; R.S.V. Pullin, T. Bhukaswan, K. common carp Cyprinus carpio on freshwater ecosystems:
Tonguthai, J.L. Maclean, Eds.; International Centre for A global perspective. Rev. Fish. Sci. Aquac., 13(3): 253-
Living Aquatic Resource Management: Manila, 290.
Philippines.
Wee, K.L. and S.W. Shu (1989). The nutritive value of boiled
Ogino, C. (1980). Requirements of carp and rainbow trout for full-fat soybean meal in pelleted feed for Nile tilapia.
essential amino acids. Bull. Jpn. Soc. Sci. Fish., 46: 171- Aquaculture, 81: 303-314.
174.
Yousif, O.M., G.A. Alhadrami and M. Pessarsaki
Olvera-Novoa, M.A., G.S. Campos, M.G. Sabido and C.A.M. (1994). Evaluation of dehydrated alfa and salt bush atriplex
Palacios (1990). The use of alfalfa leaf protein concentrates leaves in diets for Tilapia. Aquaculture, 126: 341-347.