Basrah Journal of Veterinary Research,Vol.19, No.3, 2020. Proceeding of the 17th International Conference. College of
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EFFECT OF Chlorella MICROALGAE AND GERMINATED BARLEY
POWDER ON PERFORMANCE, SOME HEALTH INDICES, AND
MEAT HYGIENE PARAMETERS OF COMMON CARP (Cyprinus
carpio)
Nasreen Mohialddin Abdulrahman*, Havan Dwud Sleman *, Derin Omer
Muhammed Ramzi*, Hevar Araz Hama-Salih**
*College of veterinary medicine, University of Sulaimani, Sulaimaniya, Iraq.
**Municipality of sulaimani, directorate of gardeners, Sulaimaniya, Iraq.
Corresponding author: nasreen.abdulrahman@univsul.edu.iq
Keywords: Chlorella, germinated barley, Cyprinus carpi.
,
ABSTRACT
This experiment was done to assess the effect of Chlorella, as a source of protein, and
germinated barley powder, as a source of natural prebiotic, on the performance of common
carp. The experiment was conducted by using one-way ANOVA (analysis of variance) with a
completely randomized design (CRD). Higher significantly in T2 with 5% Chlorella and T5
(20% germinated barley). FCR recorded significant differences among treatment as compared
to T5 (20% germinated barley), and the opposite was observed in FER. T5 with 20%
germinated barley was differing significantly in each of the spleen somatic and kidney
somatic indices. Adding germinated barley leads to enhanced intestine weight index in T4 and
T5. The parameters were hepatosomatic and gill somatic indices, condition factor, fish weight
without viscera, fish weight without viscera and head, lipids, ash and moisture, organoleptic
evaluation of mean juiciness, flavor, color, and complete acceptance. We conclude that the
use of Chlorella and germinated barley potentially enhances the growth performance and fish
Basrah Journal of Veterinary Research,Vol.19, No.3, 2020. Proceeding of the 17th International Conference. College of
Veterinary Medicine. University of Basrah. Iraq.
219
meat quality. We recommend using both plants in earthen ponds to rely on using natural
products in fish feeding.
INTRODUCTION
The fastest rising food-production technology in the world is aquaculture (1,2) stated
that Fish and other aquatic products rich in protein, essential fatty acids, vitamins, and
minerals play an significant role in international efforts to reduce malnutrition and hunger.
(3,4) reported that the microalgae have an excellent nutrient composition and are free of
poisons that could pose a threat through transferring up the food chain. In fish farming, the
chief microalgae utilization is associated with their use for feed purposes (4).
According to (4,5) around 30 per cent of the world's algae production can be used for feeding
stuffs. However, (4,6) demonstrated that microalgae are primarily utilized for larval fish,
crustaceans, and mollusks.
When Chlorella is cultivated under favorable environmental conditions, the biomass
of Chlorella might comprise 12–15% lipids and 10–15% carbohydrates. The C16 and C18
fatty acid groups such as C16:0, C16:2, C18:1, C18:2, and C18:3 are the main lipids found in
Chlorella,Moreover, chlorophyll, in conjunction with a variety of carotenoids such as β-
carotene, neoxanthin, violaxanthin, lutein, zeaxanthin, and antheraxanthin are also included in
Chlorella. The amplest carotenoid in Chlorella cells is lutein, which may amass around
0.45% of the dry weight of cells (7).
Prebiotics can reduce the risk of some illnesses and improve health. Comprehending
the connection between diet and consumer health has increased demand for real knowledge on
prebiotics in recent years. Prebiotics has become a rising sector on the world market, boosted
by technical advances, new merchandise production and the increasing number of healthconscious
consumers (8).
Prebiotics have a long history of safe use and are proven to support human health,
including increased mineral bioavailability, immune system control, gastrointestinal (GI)
infection prevention, inflammatory conditions, metabolic disorder control and cancer risk
reduction(8) Prebiotics are indigestible substances which allow specific changes in the
Basrah Journal of Veterinary Research,Vol.19, No.3, 2020. Proceeding of the 17th International Conference. College of
Veterinary Medicine. University of Basrah. Iraq.
220
composition or activity of gastrointestinal microbiota or both, which have a positive effect on
the nutrition and health status of the host (9) Prebiotics play an important role in host health
when developed by beneficial microbiota in the intestine (10,11).
Barley is widely used in both existing and new end-use applications. Much of the
barley produced is fed to animals in most countries, in particular cattle and pigs. There is
minimal use of the barley for human food. Recent developments in the use of varieties of
barley, high in dietary fiber, have however been established. The successful use of high-value
products from barley is to manufacture malt as a raw material for the ferm (6). (12) have
reported a germinated barley stock rich in arabinoxylan to facilitate the development of
bifidobacteria in humans intestines. However, there is still evidence of a consistent clinical
advantage of prebiotics in the treatment of irritable bowel disease (IBD) in extensive,
placebo-controlled studies (13).
This research aims to compare the utilization of Chlorella, a natural source of animal
protein, and germinated barley powder, a source of prebiotic, as supplements to commercial
feed for common carp.
MATERIALS AND METHODS
Experimental fish: The study lasted 84 days on 90 common carp brought from
Peramagrun/Sulaimani/Iraq. Fish weights averaged 53.12 ± 3.34 g. Fish spread in
experimental plastic containers, pre-acclimatization laboratory and fed with industrial pellets
30 days before the real feeding trials. Compositions of feed are illustrated in Table 1, and the
feeding amount was 3.0% of body weight.
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Table 1: Composition of experimental diet
Ingredients Percentage (%)
Yellow corn 15 %
Wheat bran 15 %
Soya bean meal 48% 35 %
Animal concentrate protein 20 %
Barley 15 %
Total 100
Calculated chemical composition
Crud protein 28.06
Gross energy (kcal/kg feed) 2242.7
Plastic tanks with a capacity of 70.0 L were used in this trial. Proper continuous
aeration by means of air compressors added to each tank (Hailea ACO-318, 45.0 watts power,
70.0 L/min airflow). Each replicate was stocked with six fish. The experimental trial
represented five treatments, assigned T1 through T5, with three replicates and six fish per
replicate. Group T1 was given a standard diet and served as the control. Groups T2 and T3 fed
with 5 g/kg and 10 g/kg Chlorella, respectively. Groups T4 and T5 were given a diet
supplemented with 10 g/kg and 20 g/kg germinated barley, respectively. Experimental diets
included typical ingredients found in the city markets of Sulaimani, enriched with Chlorella,
and powdered barley.
Growth and feed utilization parameters: All fish were weighed bimonthly. The feed
consumption per replicate was modified every two weeks by the biomass obtained. Weight
gained by fish (g/fish) was calculated as fish weight (g) at the end of the experiment divided
by fish weight (g) at the beginning of the experiment. Daily weight gain of fish (g/day) was
calculated as fish weight gain divided by the duration of the experiment (84 days). The
relative growth rate (RGR %) was estimated as weight gain/initial weight × 100. Moreover,
the specific growth rate (SGR) was computed according to the formula [(ln final fish weight –
Ln initial fish weight)/duration of the experiment] × 100 (14).
Feed conversion ratio (FCR) was estimated by dividing the total ingested feed (g) by
the total wet weight gain (g),Feed efficiency ratio (FER) was calculated by dividing total
Basrah Journal of Veterinary Research,Vol.19, No.3, 2020. Proceeding of the 17th International Conference. College of
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222
weight gain (g) by total ingested feed (g), Further, the protein efficiency ratio (PER) was
computed as total wet weight gain (g/fish)/amount of protein fed (g/fish) (15).
Fish weight index% was estimated by dividing fish weight without viscera (g) by fish
weight (g) and multiplying the result by 100. Finally, meat weight index% was calculated as
fish weight without viscera and head (g) / fish weight (g) × 100.
Proximate composition: All parts of fish were used to analyze the muscles chemically,
including percentages of moisture, protein, ether extract, and ash contents (16).
Moisture was measured by putting fish samples in an oven at 105 °C for 24 hours.
Then, the samples were weighed and put again in the oven for two hours and weighed till a
stable weight obtained. Fat extraction was made by Soxhlet using organic hexane CH3
(CH2)4CH3 by heating for 16 hours to determine the crude fat content.
The crude protein content of fish determined by Micro-Kjeldahl equipment, which was
done by measuring the total nitrogen content in the samples after digestion with H2SO4 and
Perchloric acid (HCIO4). Later, one part of each of potassium sulfate, copper sulfate, and
titanium dioxide, were added and the mixer was left for two hours. Distillation was conducted
by boric acid and titration with HCl, and the number obtained was multiplied by 6.25 for
nitrogen extraction of each sample. The percentage of ash content was calculated by burning
the samples in a muffle furnace at ٥٩٥ °C, till a stable weight was obtained, and weighing the
remainder (16).
Sensory evaluation: Seven experienced evaluators tasted the fish fillets and each member
filled the sensory evaluation Table as 5 = extremely like; 4 = like; 3 = neither like nor dislike;
2 = dislike; 1 = extremely dislike. The fish fillet specimens were put in aluminum containers
and cooked for 15 min in a preheated oven at 200°C.
Statistical analysis: a completely randomized design (CRD) was used, and a comparison
between the groups was made using a one-way analysis of variance (ANOVA), followed by
Duncan’s post hoc. A probability level lower than 0.05 was considered statistically
significant.
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RESULTS AND DISCUSSION
Table (2) explain the effect of adding Chlorella and germinated barley powder on the
performance of common carp, which was significantly higher in T2, with 5% Chlorella, and
T5, 20% germinated barley. There were no variations in the overall consumed diet in all
treatments. FCR recorded significant differences among treatments as compared to T5, and
the opposite was noticed in FER.
Table 2: Effect of microalgae Chlorella and germinated barely powder on common carp
Cyprinus carpio performance after a rearing experiment for 84 days
Treatments
Initial
Wt.
Final
wt.
Wt. Gain DGR % SGR RWG FCR FER
T1
Control
55.75 a 60.45 c 4.699 c 0.098 c 174.485 b 7.712 d 7.954 a 0.159 b
T2
5 % Chlorella
53.25 a 71.46 a 18.210 a 0.379 a 181.791 a 25.422 a 2.247 b 0.451 a
T3
10 % Chlorella
53.49 a 60.16 c 6.671 c 0.139 c 174.303 b 10.994 cd 6.471 ab 0.169 b
T4
20 % germinated
barely
54.17 a 64.71 b
10.536
bc
0.220 bc 177.469 ab 16.224 bc 3.690 ab 0.278 ab
T5
20 % germinated
barely
55.53 a 71.55 a
16.015
ab
0.334 ab 181.666 a 21.880 ab 2.786 b 0.465 a
Pr > F 0.002 0.052 0.004 0.004 0.007 0.002 0.055 0.048
The addition of germinated barley to fish feed has led to increased weight index of the
intestine in T4 and T5, as recorded in Table (3). The results of the fish condition factor did not
show significant differences.
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Table 3: Effect of microalgae Chlorella and germinated barely powder on common carp
Cyprinus carpio biological indices after a rearing experiment for 84 days
Treatments
Intestine weight
index
Intestine length
index
Fish Weight
without Viscera
Fish Weight
without Viscera
& Head
Condition
factor
T1
Control
2.193 ab 43.764 a 85.842 a 55.382 a 1.443 a
T2
5 % Chlorella
1.863 b 31.705 c 84.355 a 55.814 a 1.489 a
T3
10 % Chlorella
2.228 ab 41.491 ab 83.681 a 54.320 a 1.444 a
T4
20 % germinated barely
2.517 a 40.318 ab 83.469 a 53.867 a 1.530 a
T5
20 % germinated barely
2.514 a 33.057 bc 82.258 a 55.044 a 1.600 a
Pr > F 0.158 0.016 0.310 0.934 0.258
No significant differences were seen in each of the fish's weight without viscera and
fish’s weight without viscera and head. Group T3, which administered 10 g/kg Chlorella, had
more protein, while no significant differences were seen in the percentages of lipids, ash, and
moisture (Table 4).
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Table 4: Effect of microalgae Chlorella and germinated barely powder on common carp
Cyprinus carpio proximate analyses after a rearing experiment for 84 days
Treatments Protein % Lipids % Ash % Moisture %
T1
Control
19.312 6.351 1.443 71.434
T2
5 % Chlorella
19.954 6.388 1.764 71.642
T3
10 % Chlorella
20.043 6.569 1.783 71.876
T4
20 % germinated barely
19.573 6.669 1.891 71.998
T5
20 % germinated barely
19.826 6.760 1.921 72.032
Pr > F 0.004 0.004 0.002 0.007
No significant differences were observed in the organoleptic evaluation of juiciness,
flavor, color, and complete acceptance (Table 5).
Table 5: Effect of microalgae Chlorella and germinated barely powder on common carp
Cyprinus carpio organoleptic evaluation after a rearing experiment for 84 days
Treatments Juiciness Flavor Color Complete acceptable
T1
Control 4.000 a 3.876a 4.600 a 3.780 a
T2
5 % Chlorella 4.000 a 4.544 a 4.560 a 3.980 a
T3
10 % Chlorella 3.855 a 3.766 a 3.955 a 3.655 a
T4
20 % germinated barely 3.766 a 3.765 a 3.860 a 3.455 a
T5
20 % germinated barely 3.456 a 3.555 a 3.650 a 3.655 a
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Plant sources of proteins are considered a better replacement of fishmeal in aquafeed
but are usually missing amino and fatty acids. The microalga is rich in amino acids including
methionine, lysine and alanine, C. vulgaris is a perfect source of protein in catfish that may
replace fishmeal in catfish diets, and C. vulgaris meal was entirely agreeable as a feed item
and ensured that fish can utilize it well. Diet approval with high SGR lowers the cost of
aquafeeds as the planting of the algae is cheap, and it can be cost-saving compared to fishmeal
(17). Algae may be a potential source of protein in a number of fish species, for instance when
the Spirulina algae replaced fishmeal (18).
(19) observed that adding algae in fish diets enhances growth. The supplementation of
a high level of C. vulgaris (25%) may lead to a preferable growth rate of Clarias gariepinus
as compared to fishmeal, and 10.0% Spirulina had a potential increase in most studied traits
(20).
(21) observed that 50% addition of Chlorella to Macrobrachium rosenbergii enhances
SGR, weight gain, survival rate, and feed efficiency ratio. (22) concluded that mixing of
Chlorella with rapeseed powder could fully replace fishmeal in diets of crussian carp.
Chlorella is rich in amino acids like alanine, leucine, aspartic acid, serine, and glycine, and
these may be responsible for more than 50% of the total content of the Chlorella spp. (23).
As PCR increased, the FCR reduced with the addition of algal meal. This outcome
may be due to the collective effects of both amino and fatty acids on the nutrient digestibility.
(19) observed that adding algae meal improved FCR, while (20) noted that FCR was lower
when the fish had been fed on artificial diets with various algal meal rates. (24) concluded that
feeding on a diet supplemented with 20% S. platensis enhanced performance, proximate
analyses, and body weight gain of Nile tilapia. A linear weight enhancement was noted when
Chlorella was added, which may be related to lipids enhanced in fish. Moreover, the diets in
the present study had high addition levels of carbohydrate, well utilized by carp, but they
converted them to fat and stored in the fish.
Using algae in the diet of C. gariepinus improved the utilization of carbohydrates. The
addition of carbohydrate in the diets of catfish usually leads to an increased HIS, and this had
been associated with high carbohydrate diets (20, 25). These findings agree with the results of
(22), who noted that Chlorella and rapeseed meal could completely replace fishmeal. The
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reason may be organoleptic as the fish were grazing on the algae on their own before the
experiment. It is a normal behavior in fish to graze on algae while carnivores may reject the
algae powder inclusion.
The addition of algae, especially Chlorella, may be nutritionally favorable, leading to
an increased total content of fats, fatty acids, and polyunsaturated fatty acids (PUFAs) in
groups of fish fed the green algae. In some length / weight and nutritional values, the
inclusion of 10 percent algal powder in the complete carp diet formulas showed significant
variances. Adding toxic cyanobacteria reported an impairment of the fish's physiological
state. Most of the monitored values were better in the green algae groups compared to the
control fish group. Algae from a sewage water treatment lagoon were used as an additive to
fish feed to increase the levels of polyunsaturated fatty acids (PUFAs) of lipids in fish (26).
(27) showed that the average weight of carcass, crude protein, fat, and ash content
were significantly higher (P < 0.05) when fishmeal was supplemented with 20.0% Spirulina
in the diet of common carp. The results of (28) indicated that a significant increase (P < 0.05)
was observed in total weight gain, daily weight gain, relative growth rate, and specific growth
rate after the supplementation of feed with 7.5 g/kg. The significant differences (P < 0.05)
were an elevated feed conversion ratio, feed efficiency ratio, and protein efficiency ratio.
The addition of 5.0 g/kg germinated barley to aquafeed caused a significant increase
(P < 0.05) in growth performance and some blood parameters (29). However, hydroponic
germination (5.0 g) and germinated barley (2.5 g) significantly increased on protein efficiency
ratio and the intestine length index, which is following the present results.
The results in the present study agree with those reported by (30). When the diet was
supplemented with 5.0 g/kg Spirulina powder to feed common carp (C. carpio) fingerlings for
42 days, the results showed increased weight gain, and the daily, relative, and specific growth
rates. Furthermore, the feed efficiency ratio was significantly higher with adding 5.0 g and 3.0
g Spirulina powder, compared to the control group. The food conversion ratio in the control
group was significantly higher than other treatments.
The study of (31) showed that feeding algae as a feed additive to fish remarkably
increased the fish weight without viscera, and weight without viscera and head, and all these
agree with the recent results.
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Adding 7.5 g/kg Chlorella powder in the common carp fingerlings’ diet resulted in a
better weight gain (35.1 g) and daily growth rate (0.4 g) than other treatments. Moreover, the
relative growth rate (88.9) and specific growth rate (0.4) were higher than other treatments
(32). The food conversion ratio (0.6) was significantly higher when the diet was supplemented
with 2.5g/kg Chlorella powder to common carp fingerlings’ feed for 105 days. The food
efficiency ratio (2.4) and protein efficiency ratio (829.3) in the group fed 7.5 g/kg diet of
Chlorella spp. was significantly higher than other treatments.The inclusion of Chlorella and
Daphnia to fish diet in the study of (33) changed the growth performance in which it
increased the daily and final weight gain, and specific and relative growth rates. Utilization of
Chlorella and germinated barley caused a reduction in feed conversion ratio, which has a
substantial role in determining the cost of aquaculture. Dietary Chlorella and germinated
barley escalated the feed efficiency ratio and reduced feed conversion ratio. Following the
findings of chemical composition, we conclude that a high proportion of Chlorella and
germinated barley significantly affects the protein and fat ratios of fish.
Acknowledgments: The authors have unique gratitude to Dr. Hemn Nurallddin, for doing the
statistical analyses of the results, and to Mr. Amanj Baiz for helping us.
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