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© 2020 JETIR November 2020, Volume 7, Issue 11 www.jetir.org (ISSN-2349-5162)
JETIR2011273
Journal of Emerging Technologies and Innovative Research (JETIR) www.jetir.org
976
Effect of Muskmelon, Cucumis melo and Pears, Pyrus
communis extract on growth, haematology, immune
response and disease resistance in Labeo rohita
against Pseudomonas aeruginosa
Krishnasamy Pavithra1, Gopalan Vijayadurga2, Devakumar Dinesh1, Venkatachalam Ramasubramanian1*
1Division of Aquatic Biotechnology and Live Feed culture, Department of Zoology, School of Life Sciences,
Bharathiar University, Coimbatore 641046, Tamil Nadu, India.
2CAS in Marine Biology, Annamalai University, Parangipettai -608502, Tamil Nadu, India.
* Corresponding Author
1*Professor, Division of Aquatic Biotechnology and Live Feed culture, Bharathiar University, Coimbatore 641046,
Tamil Nadu
Abstract
The 45 days feeding trial was carried out to assess the effects on the Labeo rohita fish growth, biochemical
analysis, hematological parameters, disease resistance to against the fish pathogen Pseudomonas aeruginosa. The
Immune response of L. rohita when fed with using fruits methanol extraction of Cucumis melo (CM) and Pyrus
communis (PC) compared to a control diets. The fishes were randomly stocked into five experimental groups, and
diets containing different concentrations for control 0% (basal diets), 1% (CM and PC), 2% (CM and PC), were
fed to triplicate groups of fish (average weight; 2.60 ± 0.02) for 45 days. At the end of the feeding trail, test fish
were intraperitoneal injected with a pathogen, Pseudomonas aeruginosa. Resulted in significant (P < 0.05) higher
weight gain and feed intake were noticed in groups of fish fed 2% CM-treated diets compared to PM treated diets.
At the same time, none of the biochemical and hematological parameters was significantly affected. After 45 days
bacterial challenge, labeo rohita fed 2% Cucumis melo had significantly higher survival rate compare to the PM
and control groups. Moreover, Immune parameters such as, phagocytic activities were most significantly enhanced
in 2% CM especially after a bacterial challenge. This study revealed that dietary Cucumis melo at 2% could
promote the growth, Immune system, Cost effective, eco friendly alternative antibiotics and improved survival of
labeo rohita compared to PC.
Keywords: labeo rohita; Cucumis melo; Pyrus communis; Bacterial pathogen
I. Introduction
Aquaculture is currently fast-growing food-producing trade worldwide. Large-scale aquaculture
production is conquered by freshwater fish species (56.4%), especially carp species (71.9%) (FAO, 2012). Labeo
rohita (rohu) is most important Indian major fish carps. L. rohita one of the most popular and delicious freshwater
fish grow developed in India and other near closest countries. Moreover, it has been higher enlargement growth
potential and important nutritional source of protein and giant human preference (Memon et al., 2010). Escalation
© 2020 JETIR November 2020, Volume 7, Issue 11 www.jetir.org (ISSN-2349-5162)
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of aquaculture operations, which is now universal trend for commercial aquaculture (Msangi et al., 2013), is
generally accompanied by stress leading fish growth fall, immune suppression, susceptibility to contagious
diseases and causing major monetary loss to the farmers (Abdelsalam et al., 2017; Food and agriculture
organization, 2016; Mona et al., 2015; Syahidah et al., 2015).
In addition, Fishes are always vulnerable to a wide range of pathogenic bacterial strains (Banerjee et al.,
2016). Bacterial diseases in fish are the major challenges that obstruct the production and affect the country
financial system. Pesudomonas aeruginosa is a gram-negative, rod shaped bacterium belonging to the family
Psedomonadaceae. This species very important modifiable and environmental opportunistic pathogen, capacity of
surviving in environmental sphere, including aquatic resources (Abdullahi et al., 2013). (Thomas et al., 2014)
which causes skin darkness, abdominal ascites, exopthalmia, ulcerative syndrome, dropsy, gill rot, destruction of
the primary and secondary lamella in fishes. Therefore, many problems have been most part addressed with the use
of antibiotics that leads to the drug-resistant to pathogens, the impact of released antibiotics on the atmosphere and
food safety concerns (Upadhaya et al., 2017).
Recently, the use of immunostimulants in the fish culture developed industry has been important in
dropping antibiotic use and profitable losses (Citarasu, 2010; Hoseinifar et al., 2016). There is upward in the
substitution of chemical drugs with herbal medicine and algal extracts, due to their possible health benefits with
inadequate side effects (Peddie, Zou & Secombes 2002; Najafian & Babji 2012; Faggio et al., 2015).Furthermore,
Plants with traditional medicinal usage have been identified as potential feed additives for increase resistance to
pathogens, improved feed efficiency and growth performance subsequently (Setufe et al., 2018). Some products of
herbal plants have been play vital role in host innate immunity and activation of macrophages, neutrophils, and
lymphocytes at infection sites for pathogen elimination, promotes hematological and biochemical performance,
enhance the fish growth and also protects from the diseases (Johnson and Banerji, 2007).
Now days, fruits also have been studied due to their numerous and important health benefits such as free
radical scavengers, anti-oxidants, immunostimulants, anti-inflammatory, anti-cancer and anti-microbial activity
(Dragsted, Strube and Larsen, 1993). Cucumis melo fruits are a luscious, delicious fruit of the Cucurbitaceae
family. It is civilized in all the temperature regions of the worldwide due to its good adaptation to soil and climate.
A study previously done by Vouldoukis et al. (2004) found that the muskmelon pulp extract contained anti-
inflammatory, high antioxidant properties posses’ useful medicinal properties activities of anti-platelet, anti ulcer,
anti-cancer, anti-microbial, hepatoprotective, anti-diurectic, anti-diabetic, anti-helmintic and anti-fertility activity.
Another fruits, Pyrus communis belongs to Rosaceae family. P. communis fruits used as a conventional
folk therapy for relieving alcoholism, eliminating constipation, relieving cough, and moistening the lung for more
than 2000 years in Asian countries, particularly in China, Japan, and Korea. Further, bioactive compounds such as
polyphenols are an important group of secondary metabolities distributed in the plant kingdom and these active
compounds are usually more rigorous in the peel than in the fruit flesh (Andreotti, Costa & Treutter, 2006;
Kolniak-Ostek, 2016). Pear fruits properties of antioxidant and anti-inflammatory affects their products (Li et al.,
2014).
Therefore, the fruits compounds properties against the fish pathogen and did not affect the fish growth.
This study is investigated for assessing the effect of C. melo and P. communis extract on growth performance,
biochemical analysis, hematological parameters, immune response of labeo rohita fingerlings and its Susceptility
to pathogenic Pesudomonas aeruginosa.
II. Materials and Methods
2.1 Sample collection (fruits) and preparation
The Musk melon and Pears were collected from the local Super Market Vadavalli Coimbatore, Tamilnadu.
The fruits were washed to remove the any dirt adhering to the surface and peeled the outer layer of the fruits. The
separation of the pulp with seed was done manually. After the separation the pulp is pounded into small pieces and
then dried. The pieces were shade dried. They were Powdered by mixer grinder and stored at -20˚c for further use.
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2.2 Extraction of Muskmelon (Cucumis melo) and Pears (Pyrus communis)
The fruit powder (100g) for each fruit was packed into the extraction chamber of the soxhelt extractor.
While a solvent (methanol 99%) 250 ml was poured into the round bottom flask of the extractor. The whole set up
was mounted on a heating mantle at 65˚c and allowed to reflux for about 16 hours. The extract was filtered (to
remove impurities) and evaporated using a rotary evaporator to isolate the free flow lipid from the solvent. The
residues obtained after evaporation of methanol was kept in sterilized screw cap glass container for further use.
2.3 Collection and Maintenance of Experimental Animal
In the present study Labeo rohita fingerlings (2.60±0.02) were used as a experimental organism. Healthy
and diseases free fingerlings of Labeo rohita fishes were purchased from Pandian fish seed farm Dindigul,
Tamilnadu, India. Specimens were acclimatization fish 2 weeks. The investigational animals were fed with rice
bran for maintenance. Thereafter, specimens were randomly distributed into five poly vinyl circulating troughs
(75L) at a density of 20 fish per tank. Fish were hand-fed twice a day (09:00 and 15:00) as described elsewhere
(Roosta, et al. 2014). Treatment was carried out under static aerated water conditions with 50% water change
every day. For this purpose an 45-days feeding trail was also conducted under the same experimental setup in
order to determine the appropriate ration size of the fish by feeding fish at the rate of 0%, 1%, 2% BW/day. The
uneaten feed was collected after active feeding approximately for 40 min with the help of siphoning pipe and
collection tubes. The collected feed was then oven-dried at 100 ͦ C to calculate the final feed conversion ratio
(FCR). No feed was offered to the fish on the day of weekly measurement. At the end of the experimental trail,
desired numbers of fish were randomly sacrificed for the assessment of whole body composition.
2.4 Diets analysis
Diets were analysed for proximate composition using method described by the Association of Official
Analytical Chemists (AOAC, 2005) Table 1.
Table 1 Proximate and chemical composition of the experimental diets (g/kg)
Ingredient
Control (g)
T1 (CM-1%)g
T2 (CM-2%)g
T3 (PC-1%)g
T4 (PC-2%)g
Fish meal
30
30
30
30
30
Soybean meal
20
20
20
20
20
Rice bran
20
20
20
20
20
Groundnut oil cake
10
10
10
10
10
Wheat flour
5
5
5
5
5
Tapioca flour
5
5
3
5
3
Egg albumin
3
3
3
3
3
Cod liver oil
3
3
3
3
3
Vitamins & mineral mixture
4
2
2
2
2
Fruit extract
0
2
4
2
4
Total
100
100
100
100
100
Proximate Composition %
Moisture Content
9.70
9.25
8.75
9.50
9.10
Crude fibre
3.60
3.85
4.22
3.90
4.42
Ash
4.88
5.20
5.93
5.18
5.46
Note; Diets were prepared with locally available feed ingredients.
Fishmeal (150 g kg-1) and soybean meal (100 g kg-1) were used as protein sources; rice bran (100 g kg-1) ; groundnut oil cake (50 g kg-1)
wheat flour (25 g kg-1) Tapioca flour (25g) were used as carbohydrate sources; cod liver oil (15 g kg-1) was used as lipid source; Binding
agent egg albumin (15 g kg-1) was also added; composition of vitamins mineral mixture tablets sadamindia Ltd, Mumbai, India (vitamin
A, cholecalciferol, thiamine mononitrate, riboflavin, pyridoxine hydrochloride, cyanocobalamin, nicotinamide, calcium pantothenate,
© 2020 JETIR November 2020, Volume 7, Issue 11 www.jetir.org (ISSN-2349-5162)
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ascorbic acid, α-tocophenyl acetate, biotin (20 g); mineral mixture (cu,zn free) sodium molybdate dehydrate, sodium borate, black oxide
of iron, red oxide of iron, magnesium sulphate, manganese sulphate, cellulose (20 g kg-1) was also added.
2.5 Growth performance
Before measuring the growth performance of L. rohita were starved for 24 hrs. Eight fish were randomly
selected from each experimental groups and transfer on the tray. Total weight gain, Specific growth rate, and feed
conversion ratio were calculated (castell J & Tiews K, 1980).
Various parameters as follows:
Total gain (g fish-1) = WT-WI, where as WT is a final weight, WI is a initial weight
The specific growth rate (SGR-% day-1) = 100× (In WT-In WT)/duration/day
The feed conversion ratio (FCR) = total feed intake (g)/total gain (g).
Survival percentage = Number of fish in each group remaining after the 45- days period/ initial number of fish
× 100
2.6. Biochemical and Haematological analyses of Labio rohita fed experimental diets
At the end of the 45 –day feeding trail, 10 fish per treatment were randomly caught using hand net and blood
was drawn from the caudal vein region of fishes by using a hypodermic syringe that was previously rinsed with
EDTA an anti-coagulant. The collected blood was diluted with EDTA (2mg⁄ml). The hemoglobin concentration of
blood was estimated by the method using Sahli’s haemoglobinometer (Put, 1923).
White blood cells were determined using a Neubauer haemocytometer as described by Kaplow (1955).
Erythrocytes indices in the blood were determined according to Jain (1986). The protein content in different
samples was estimated by employing the
Folin-Ciocalteau method of (Lowry et al., 1951). Finally, colour developed was read at 620 nm and with UV
spectrophotometer. Standards were also run simultaneously with OD value the total protein content of the sample
was calculated. The method of, Rob (1955) was employed to 500 mg of the sample was homogenized with 100ml
of distilled water and centrifuged for the 5 minutes at 4000 rpm. The supernatant was used for the estimation of
carbohydrate. The amounts of carbohydrate present in the sample were calculated. The method of Folch et al.
(1957) was employed to extract the 500mg sample was homogenized in 2ml of chloroform: methanol, which is
centrifuged at 3000 rpm for 10 minutes. The process is repeated thrice; the supernatant was pooled and
concentrated at 40- 45ºC in flask vacuum evaporator. The total lipid concentration of the sample was calculated
from the standard graph and expressed as mg ⁄ml wet tissue.
2.7 Challenging study
Bacterial strain of P. aeruginosa was purchased from PSG Medical College, Coimbatore. The bacteria
cultured in nutrient broth at 37˚c for 24 hours. The bacteria culture was adjusted to 1 × 106 CFU/ ml by serial
dilution. Twenty fish randomly selected from each experimental group and divided into two groups each of 6 fish.
The first group was intraperitoneally injected with 0.2 ml PBS containing 1 × 106 CFU/ ml virulent of P.
aeruginosa (Kocour et al., 2005). The second group was intraperitoneally injected with 0.2ml of saline solution as
a control. Before infection, fish were starved for 24 hrs. All fish were kept under observation for 15 days to record
the daily mortalities and abnormal clinical signs. Relative percentage survivals (RPS) among the challenge fish
were determined using following equation (Misra et al., 2006).
RPS% = Number of surviving fish after challenge/ Number of fish injected with bacteria ×100
2.8 Phagocytic activity
Fishes were immobilized by a sharp blow to the cranium, and about 2 ml of blood collected from the
caudal vein using a heparin-treated,150 units ml-1 syringe, 0.1 ml of Staphylococcus aureus, 1 × 108 ml-1
suspended in PBS was added to 0.1 ml of blood in the microtiter well. The amalgamation was incubated at room
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980
temperature for 20 min and mixed thoroughly using a pipette to ensure contact of the bacteria and leukocytes. Fifty
microliters were placed on a glass slide to make a smear similar to the method of making a blood smear. The slide
was air dried, fixed with 95% ethanol, re-dried, and then stained with 0.15% safranin solution for 10 min. The
phagocytic cells were observed and engulfed bacteria were counted for calculation of phagocytic index. The
percent phagocytosis was calculated according to the following after counting at least 100 phagocytic cells either
phagocytizing or not (Mamnur rashid, 2002).
PA% = Number of phagocytizing cells ×100 / Total number of phagocyte cells counted
2.9 Statistic analysis
Results for each parameter were expressed as the mean ± SD. Data were analyzed using one-way, ANOVA
and the comparison of the mean values were done by using Duncan multiple range test using software program
SPSS version 16 for windows. Differences were considered statically significant when, P<0.05.
III. Results
3.1 Growth performance
Growth parameters of L. rohita fed on CM and PC supplemented diet are presented in Table 2. There was a
steady increase in the weight of all experimental groups during the whole experimental period. After feeding trail,
weight gain (WG %) and Specific growth rate (SGR %) of experimental fish showed significant higher (P < 0.05)
values in 2 % CM compared to the control. Fish fed with 2 % CM supplementation had the optimum fish growth
producing the lowest FCR value (P < 0.05) and another parameter of fish survival rate (%) attained the two fruits
using cucumis melo and pyrus communis for 45 days. In this results analysed 100% survival rate were observed by
using CM (2%) treated when compared other treated groups.
Table 2 Growth parameters and feed utilization of Labeo rohita fed on diets containing different cucumis melo and pyrus
communis fruits extract for 45 days
Note; Regressions were measured significant at p < 0.05.WG, Weight Gain; SGR, Specific growth rate; FCR, Feed conversion ratio; Data
were expressed as Mean ±SD, n = 3 replicates; a,b,c ,Mean values in the same row with different superscripts differ significantly (p <
0.05).
3.2 Hematological Parameters
The hematological profiles of labeo rohita fed C. melo and P. communis extract‐based diets are presented
in Table 3 (Fig.1). There was a significant increase (P < 0.05) in the recorded values for Hb, RBC in the 2 %
cucumis melo treated groups when compared with the control. Highest values of Hb concentration (10.09), RBC
(2.56) were recorded for T2 while control had the least values for these parameters. The C.melo and P communis
S. No
Control
T1 (CM-1%)
T2 (CM-2%)
T3 (PC-1%)
T4 (PC-2%)
Initial weight (g)
2.6±0.08
2.76±0.18
2.99±0.08
2.83±0.26
2.73±0.16
Final weight (g)
6.93±0.04
8.57±0.01
9.71±0.02
8.89±0.004
6.29±0.01
WG
4.33±0.23d
5.86±0.12c
7.26± 0.20a
6.066±0.49bc
6.63±0.30ab
FCR
2.73±0.12a
2.3±0.21b
1.68±0.08d
2.06±0.16bc
1.8±0.08cd
SGR
1.42±0.05d
2.13±0.26c
3.2±0.35a
1.98±0.23bc
2.58±0.18b
Survival Rate %
83.5±1.22d
92±1.63c
100±0.0a
95.6±0.94b
98±0.81ab
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fruits extract using 1% and 2% separately treated groups over entire feeding period as compared to the control. The
WBC level was observed in all treated groups in particular C.melo- 2% treated was 14.25 significantly decreased
when compared to control.
3.3 Biochemical parameters
Protein content in muscle of the fish was significantly increased, P<0.05 in all treated groups with
muskmelon and pears on days 45 compared to the control group. Table 3 shown highest protein level 17.3 in C.
melo (CM) 2 % feed when compared to P. communis (PC) 2% protein level 16.8 and control 14.6 (Fig.1). The
carbohydrate level of 2% muskmelon, 2% pears treated group and control. This result shows the carbohydrate
level increased 2.56 which treated muskmelon 2% when compared to the pears and control. Similarly biochemical
parameters the lipid level was decreased by treated Cucumis melo when compared to other treated group.
Table 3 Haematological serum biochemical parameters of Labeo rohita fed diet containing different cucumis melo and pyrus
communis fruits extract for 45 days
Note; Regressions were measured significant at p < 0.05.HB, Haemoglobin; RBC, Red Blood Cells; WBC, White
Blood Cells. Data were expressed as Mean ±SD, n = 3 replicates; a,b,c Mean values in the same row with different superscripts differ
significantly (p < 0.05).
Figure 1: Haematological serum biochemical parameters of Labeo rohita fed diet containing Treatments
0
2
4
6
8
10
12
14
16
18
20
Treament (%)
Haematological serum biochemical parameters
Control
T1 (CM-1%)
T2 (CM-2%)
T3 (PC-1%)
T4 (PC-2%)
S. No
Control
T1 (CM-1%)
T2 (CM-2%)
T3 (PC-1%)
T4 (PC-2%)
HB
8.23±0.12d
9.23±0.20bc
10.09±0.285a
8.69±0.42cd
9.61±0.10ab
RBC
1.33±0.12d
1.63±0.04c
2.56±0.16a
1.53±0.28c
2.1±0.008b
WBC
18.1±0.14a
15.2±0.69b
14.25±0.23c
13.1±0.23d
13.5±0.28cd
Protein
14.6±0.12e
16.06±0.16c
17.3±0.20a
15.23±0.12d
16.8±0.21b
Carbohydrate
1.33±0.12d
1.63±0.04c
2.56±0.16a
1.53±0.28c
2.1±0.008b
Lipid
18.1±0.14a
15.2±0.69b
13.1±0.23c
14.5±0.23d
13.5±0.28cd
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3.4 Phagocytic activity
Phagocytic activity was significantly, P<0.05 different in the experimental groups over the control. Higher
phagocytic activity percentage, 34.4% in T2 muskmelon 2% feed and 25.5% in T4 pears 2% feed as compared to
control. After challenge study, there was a significant increase in all treated groups (Table 4; Fig.2).
Table 4 Phagocytic activity of L.rohita fingerlings fed with different fruits
Muskmelon
Pears
S.No
Pre-
challenge
Post-
challenge
S.No
Pre-
challenge
Post-
challenge
Control
18.39±0.02
e
18.48±0.02
d
Control
18.39±0.02
e
18.48±0.02
d
T1 (CM-1%)
20.53±0.23
c
20.6±0.22
c
T3 (PC-1%)
19.85±0.1
d
19.97±0.1c
T2 (CM-2%)
32.7±0.36
a
34.4.±0.37
a
T4 (PC-2%)
23.1±0.2
b
25.5±0.22
b
Note; Data were expressed as Mean ±SD, n = 3replicates; a,b,c Mean values in the same row with different superscripts differ
significantly (p < 0.05).
Figure. 2 Fruits extract using Phagocytic activity of L.rohita
0
5
10
15
20
25
30
35
40
Control T1 (CM-1%) T2 (CM-2%) T3 (PC-1%) T4 (PC-2%)
Muskmelon Pears
Phagocytic activity (%)
Treatment
Pre-challenge
Post-challenge
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3.5 Challenge test
The survival rate of fish challenged with P. aeruginosa (1 × 106 CFU/ ml) for 15 days after the 45‐days
feeding experiment showed the highest mortality in control (100%), while T2 recorded the least mortality (20%).
A reduced mortality of P. aeruginosa was observed in groups fed with C.melo extract compared to PC and control
(Table 5). The survival rates of the morality were used to calculate Relative Survival Percentage (RPS).
Table 5 Challenge study of labeo rohita fed diets prepared with fruits extraction
S. No
No of fish
infected
Total No of
mortality in
15 days
% of mortality
RPS
Control
10
10
100
0
T1 (CM)
10
5
50
50
T2 (CM)
10
2
20
80
T3 (PC)
10
8
80
20
T4 (PC)
10
4
40
60
Note: Relative Percentage Survival (RPS)
IV. Discussion
According to our knowledge, there is a little information about fruits uses given in the literature. Hence, we
have selected natural promoting immune stimulant using fruits while developing the fish growth, reduced
mortality those were helpful to minimizing the use of antibiotics. This study is the foremost attempt to investigate
the end result of C. melo and P. communis fruit extract as dietary immuno-stimulant on the growth performance,
hematological performance, bio-chemical analysis and disease resistance of L. rohita. At the final stages of feeding
trail, significant difference were noticed, P<0.05 increases in weight gain have been recorded at 2 % C. melo on
various treatment groups compared to P. communis and control. However, significant difference noticed SGR,
FCR was observed between control and experimental groups. Furthermore, growth profiles has been measured and
found to be superior weight gain percentage in T4, LE diet fed fish. Similarly, significant, P<0.05 SGR found in
treated diet and highest SGR found in T4, 0.4% LE diet fed fish. Falling trends were noticed in FCR as the
quantity of lapsi extract increased. Lapsi powder supplementation diets groups T7 and T8 while compared to T2
and T3 incorporated with Choerospondias axillaris, lapsi extracts found to be performing better growth than the
lapsi powder supplemented diet fed (Shyam et al., 2017). On other hand, dietary supplementation with 5% BPF,
i.e., B5 for 60 days significantly increased WG and SGR in L. rohita (Sib Sankar Giri et al., 2016).
Assessment of nutritional status, wellbeing status, and the capability for fish adaptation to the external
environment study using that Hematological and biochemical investigations are important key tools (Abdel-
Tawwab et al., 2016; Fazio et al., 2013). Hemoglobin content was increased in 2% C. melo fruit extract.
Hematological parameters provide an index of the morphological and physiological condition of fish (Fadeifared
et al., 2018). Likewise, flavonoids give out an fundamental role in obstacle oxidation of hemoglobin by diverse
factors, such as hypochlorous acid (Gebicka et al., 2012). The hemoglobin rate was good quality indicators for
oxygen distribution capacity of fish, thus manufacture it achievable to establish relationship with the oxygen
concentration obtainable in the habitat and the health condition of these fishes (Barros et al., 2002).
We revealed that L. rohita fed with 2% C. melo fruit extract supplemented diet showed significantly higher
RBC level, P<0.05 then the P. communis and other experimental period over the feeding trails. In addition, it
constitutes the transportation organization of oxygen from the lungs to tissues. It have been verified that a huge
concentration of polyphenols might boost the resistance of RBC to oxidative stress, (Youdim et al., 2000).
Whereas, prospective elucidation for the outcome was showed a better content of polyphenols, erythropoiesis in
Pomegranate juice may have been prevented RBC destruction due to reduced oxidative stress (Eirini manthou et
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al., 2017). WBC counts were significantly, P<0.05 diverse in experimental groups over entire feeding period as
while comparing the control in T0. The higher WBC counts were noticed T2, muskmelon among other
experimental groups while lower WBC counts were seen in, T0. Adewolu et al. (2008) reported that the significant
raise in the values of WBC concentration of tobacco dirt raised could be attributed to elevate in leucocyte synthesis
as a defense mechanism against the destruction of erythrocytes. Leucocytes are most plentiful cells comprising the
lymphocytes, which may function in the production of antibodies and chemical substances serve as a defense
against contagion. Moreover, that they noted the raised in MCHC, WBC, MCV and MCH. Many studies took
down the medical plants could act as immuno-stimulants increase the WBC count, Kumar et al., 2014; Baba et al.,
2016. On the contrary, Ibrahim Adeshina et al. (2019) the hematological profile of African catfish fed ECBE-
enriched diets was significantly different, P < 0.05.
Fish species are the excellent sources of superior quality protein and possess high amount of essential and
functional amino acids which regulate and take part in various metabolic pathways and with beneficial health
effects on growth, survival, development, lactation and development of an organism (Wu, 2010). Proteins have
more essential functions in all the biological activity due to enzymatic acticit, transport, storage, mechanical
conservancy, growth and cellular differentiation control (Lowry et al. 1951). Protein contented in muscle tissue of
the fish was significantly increased, P<0.05 in all treated groups with muskmelon and pears on day 45 compared to
the control group. Furthermore, amino acids bring crucial functions in cell signaling and act as regulators of gene
expression and protein phosphorylation cascade (Wu, 2010), similar trend as their values recorded in the study
follow as (Abdel-Tawwab et al., 2010; Ulukoy et al.,2017). Ibrahim Adeshina et al. (2019) have reported that total
protein, albumin was increased after the ECBE feeding when compared to the control diet.
Carbohydrate is one of the very important components of metabolism and it supplies the energy needed for
respiration and other most important processes (Hedge et al., 1962). The Lower carbohydrate level were noted in
2% C. melo fruit extract, T4 and 2% P. communis fruit extract died fed fish and the higher carbohydrate level were
noted in control. Whereas, the study confirmed that the growth of M. montanus was more influenced by the animal
source of fishes along with the nutritional source of wheat flour. The omnivorous fish M. montanus need a low
amount, 9.48% of dietary carbohydrate for its maximum growth (Raj et al., 2008). Similarly the outcome showed
that lipid level was lower at 2% C.melo fruit extract, T4 than 2% P. communis fruit extract died fed fish and
control. Total lipid content in body parts varied in different fresh water fishes studied (Swapna et al., 2010).
Earlier studies have reported that pomegranate may reduce blood lipid levels (Basu et al., 2009).
Post-challenge mortality was significantly reduced in fish fed C. melo and P. communis-containing diets. The
highest death rate was recorded in fish fed control diet while the low mortality was obtained in fish fed at 2 % C.
melo diet. Moreover, L. acidissima fruits supplementation activated the innate immunity and reduced mortality of
the fish, (ponnuraj srinivasan et al., 2015). A pathogen challenge test evaluated the effects of ECBE as an immune
stimulant have been tolerated to pathogenic bacteria of A. hydrophila augmented significantly as dietary ECBE
levels increased, shown higher fish survival other than the control fish group. These results coincide with the
results of Abdel-Tawwab (2012); Abdel-Tawwab and Abbass (2017); Abdel-Tawwab et al. (2010); Abdel-
Tawwab et al. (2018) and Adeshina et al. (2017) who noticed an improved tolerance of Nile tilapia and African cat
fish against A. hydrophila infection.
Phagocytosis activities have been documented as an very important cellular progression in the nonspecific
immune system of fish (Bergljot Magnad´ottir et al., 2006). Phagocytic activity was significantly, P<0.05 different
in the experimental groups over the control. Higher phagocytic activity were found to be 34.4% in T2,muskmelon
4% feed and 25.5% in T2,pears 2% feed as compared to the control. Furthermore, as the primary line of defence
various peptides such as lysozyme, antibodies, and complement factors inhibit the adhesion and colonization of
microorganisms, leading to the prevention of infection and disease (Esteban et al., 2012).
V. CONCLUSION
Therefore, our results concludes that the C. melo and P. communis fruits methanol extract improves the
digestibility of protein in fish diets as well as to increase fish growth of Indian major carps (labeo rohita) and also
C. melo fruits extracts act as disease resistance of fish bacterial diseases and were proved to be maintain the
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Journal of Emerging Technologies and Innovative Research (JETIR) www.jetir.org
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sufficient health, prevent the high risk of diseases and fish mortality when compared to P. communis fruits
methanol extract.
VI. ACKNOWLEDGEMENT
In this manuscript. K. Pavithra is grateful to the Department of Science and Technology promotion of University
Research and Scientific Excellence, India (DST Purse -II) providing financial support.
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