J. Env. Bio-Sci., 2015: Vol.

29 (2):319-322
(319) ISSN 0973-6913 (Print), ISSN 0976-3384 (On Line)

STUDY OF HAEMATOLOGICAL PROFILE OF LABEO ROHITA (ROHU) FED WITH

TINOSPORA CORDIFOLIA (GILOY) AS NEUTRACEUTICAL
Anita1, Maneesh Kumar Dubey2, Akansha Khati3 and Pooja Kanyal4
1, 3
Department of Aquaculture, College of Fisheries, Pantnagar
2
Virology Laboratory, Directorate of Cold Water Fisheries, Bhimtal
4
Department of Fish Pathology, Cental Institute of Fisheries Education, Mumbai
[Corresponding author E-mail1: anitatamta20@gmail.com]

Received: 22-04-2015 Accepted: 02-06-2015

Present experiment was designed to study the effect of Tinospora cordifolia stem powder on haematology of Labeo rohita for
examination of immunological parameters. Fingerlings weighing average weight of about 15+0.2 were selected and kept in FRP
tanks. The fingerlings were divided into 4 treatment groups T0, T1, T2, and T3. The conventional feed ingredients; deoiled Mustard
cake, Soybean meal, Rice bran and Agrimin Forte (Vitamin-Mineral mixture) were used for preparation of feed pellets. The fingerlings
were fed with diet D0 (Control diet), D1 (0.25% incorporation of stem powder of Tinospora cordifolia), D2 (0.5% incorporation) and D3
(0.75% incorporation) @ 5% body weight for 90 days. At the end of the experiment the haematological examination showed
significant improvement in the fishes fed with the diet D3, which was incorporated with 0.75% of stem powder of Tinospora
cordifolia as compared to control and other treatments. The haematological parameters; TLC (42.682×103/µl) , TEC (3.219×106/µl,
hemoglobin concentration (11.21g%), total serum protein (9.23g/dl), albumin (1.201g/dl) and globulin (12.527g/dl) in D4 diet fed
fishes showed increase values in comparison to control and all other treatments (P<0.05), which depicts that the stem powder of
Tinospora cordifolia has very good potential in enhancing the immunological parameters in fingerlings of Labeo rohita, ultimately
enhancing the immunity of Labeo rohita fingerlings under tank culture system.

In Asia, Aquaculture sector contributes 89% of world high potassium content (0.845%) which assist in regulatory
aquaculture production, whereas, India ranks IInd with production function of nerve impulse, high chromium content of 0.006%
of 5.2% of world fish1. Fisheries is rapidly growing food useful in regulation of carbohydrate utilization, iron percent of
producing industry but recently various stress causing factors, 0.28% which is helpful in hematopoietic functions and calcium
pollutants, and intensive culture practices in this field is content of 0.131% which help in regulatory functions in blood
possessing serious threat related to the emergence of diseases. coagulation, nervous, cardiovascular and musculoskeletal
Several prophylactic measures, chemicals, vaccines and system)2.
antibiotics can be applied to control and treatment of such
Some researchers worked on potential of neutraceuticals in
occurrences but it has been observed that the emergence of
aquafeed and found that they are very cheap, effective and
antibiotic resistant organisms and limited use of vaccines made
eco-friendly 3 .Various investigators studied the oral
the researchers to get sustainable, eco-friendly, consumer
supplementation of microbial phytase in the diet of Labeo rohita
friendly and cheap nutrients which act as growth promoters
in which they found enhanced growth performance and nutrient
and as well as immunostimulators.Such nutrient supplements
digestibility4. Certain investigations have shown significant
are called as neutraceuticals.The herb used in the present
increase in growth of rohu fed with non-gelatinized corn
experiment is a potent immunostimulater in humans and is
supplemented with a-amylase at sub-optimal protein level5.
well known as "Amrita" in Ayurveda. It is declared as ayurvedic
Some investigators observed efficacy of different dietary doses
drug as per the Drugs and Cosmetic Act of India (1940). The
of Zingiber officinale powder for the immune response and
stem contains immunologically active substance
disease resistance of the Indian major carp (Catla catla)
sarabinogalactan and (1, 4)-alpha-D-glucan. Crude values for
infected by Aeromonas hydrophila6. Herbal extracts are being
food contents in Guduchi include high fibre (15.9%), protein
used as immunostimulators in not only fin fish sector but also
(4.5%-11.2%), carbohydrate (61.66%) and low fat (3.1%).
in shell fish culture units. Researchers investigated
Nutritive value of Giloy is 292.54 calories per 100 g. It also has
NAAS Rating (2016)-4.20
 STUDY OF HAEMATOLOGICAL PROFILE OF LABEO ROHITA
Table-1. Proximate composition of experimental diet
immunomodulatory effects of turmeric, Curcuma longa
(Magnoliophyta, Zingiberaceae) on Macrobrachium rosenbergii
(Crustacea, Palaemonidae) against Vibrio alginolyticus
(Proteobacteria, Vibrionaceae)7. The present study focuses
on the haematological examination of fingerlings of Labeo rohita
to study various immunological parameters.
MATERIAL AND METHODS
Acclimatization of fingerlings: Healthy and disease free
fingerlings, weighing average body weight 15 ± 0.2 gm of Labeo
rohita were selected from Fish seed hatchery, College of
Fisheries, G.B Pant University of Agriculture and Technology,
Pantnagar for the study. The fingerlings were kept in disinfected
FRP (Fiberglass Reinforced Plastics) tanks filled with
oxygenated water and were acclimatized for 1 week. During
acclimatization, the fingerlings were fed with normal diet
comprising of rice bran, deoiled mustard cake, soybean meal
and Agrimin Forte. The water of the culture unit was 50%
renewed every day and 100 % in a week.
Preparation of pelleted feed: For preparation of Control
diet (D0) rice bran, deoiled mustard cake, soybean meal were
semi cooked in pressure cooker for 15 min, cooled followed
by addition of Agrimin Forte (1%) and wheat flour as binder.
The mixture is then filled in pelletizer machine and pellets
were collected in aluminum tray. The pellets were then kept
(320)
for drying in oven at 60C for 24 hours, till the moisture content
was reduced to less than 10%. The dried pellets are packed
in air tight containers, labeled and stored in cool dry place.
Analysis of proximate composition of the experimental feed
also performed in the laboratory (Table-1).
The incorporation rate of stem powder of Tinospora cordifolia
in the diets was @ 0.25% in D1 diet, 0.5% in D2 diet and 0.75%
in D3 diet.
Experimental design: The present experiment was divided
into 4 treatment groups, each group consisting of 20 fingerlings
of Labeo rohita. The control treatment group (T0) fishes were
fed with control diet (D0) and other treatment group fishes were
fed with the diet incorporated with stem powder of Tinospora
cordifolia @0.25% in T1 group, 0.5% in T2 group and 0.75% in
T3 group.
Estimation of water quality parameters: Quality water is
very necessary for healthy and productive aquaculture. The
various water quality parameters of the tanks were regularly
monitored.
The temperature of experimental water varied from 300C to
minimum 200C. Water pH ranged from 7.00 to 7.7. The range
of total alkalinity was 90 to 110 mg/l, dissolved oxygen varied
from 6.2-8.0 mg/l during experimental period and free carbon
dioxide of the waters of all the FRP tanks ranged from 0.03 to
0.7 mg/l, which indicates that the culture medium of
experimental tank was favorable throughout the experimental
period8.
Haematological analysis: The fishes were caught individually
using scoop net. Blood was collected from caudal peduncle
(321) ANITA, DUBEY, KHATI AND KANYAL
region of the fish through syringe of 2ml in sterilized ependorf
tubes adding a pinch of anti coagulant, EDTA for estimation of
various immunological parameters. In case of estimation of
Serum from blood, anti-coagulant was not added. Total
erythrocyte count (TEC) and total leucocytecount (TLC)
estimation was carried out by using Haemocytometer with
improved Neubaurer ruling chamber (Weber & Sons England).
Concentration of haemoglobin was estimated by acid- haematin
method using Sahli's haemoglobinometer and the value is
expressed in g%. For the estimation of biochemical
parameters, Auto Span kits were used. Total serum protein
was estimated by Modified Biuret, End Point Assay, while
albumin levels by Bromocresol Green, End Point Assay and
was analyzed by using kits (Auto Span kits). Globulin level
was estimated by subtracting the value of albumin from total
serum protein.
Statistical analysis: Analysis of variance (ANOVA) was used
at 5% significance level to test significance differences between
the immunological parameters of different treatments9.
RESULTS AND DISCUSSION
Haematolagical examination of blood showed remarkable
improvement in all treatment groups (Fig.-1). The maximum
level of total leukocyte count was recorded in T3 treatment
group (42.682× 10³/µl), followed by T2 (29.05× 10³/µl), T1 (18.928
× 10³/µl) and minimum in T0 control (15.285× 10³/µl), (P<0.05).
Highest level of total erythrocyte count was also observed in
T3 group (3.219× 106/µl), followed by T2 (3.00× 106/µl), T1 (2.50
× 106/µl) and minimum in T0 control (2.125× 106/µl) (P<0.05).
T3 treatment fishes also showed significant improvement in
Hb level (11.21 g %), followed by T2 (11.012 g %), T1 (10.21 g
%) and minimum in T0 control (8.13 g %), (P<0.05). Total serum
protein level (Fig.-2) was also high in ( 9.233 g/dl) in group T3,
T2 treatment fishes TSP level (9.102 g/dl) high as compared to
control (8.557 g/dl) and T1 (8.932 g/dl) treatment (P<0.05).
The best level of albumin (1.201 g/dl) was recorded in T3 group,
T2 showed albumin level (1.00 g/dl) followed by T1 with albumin
level of (0.9 g/dl) and minimum in T0 control treatment (.809 g/
dl) (P<0.05). Comparatively high globulin level was recorded
in treatment T3 (12.527 g/dl), followed by treatment T2 (10.381
g/dl), treatment T1 (8.782 g/dl) and minimum in T1 control
treatment (6.978 g/dl) (P<0.05). As TEC is reliable indicator of
stress and TLC is considered as indicator of good health. Thus,
the highly significant increase in TEC, TLC and Hb levels in D3
incorporated diet fed fingerlings of Labeo rohita developed a
response to tolerate stress and their good health in tank culture
system indicates that the stem powder of Tinospora cordifolia
have good potential as anti-stress agent and immunostimulating
effect on fingerlings of Labeo rohita.
Some researchers investigated that administration of ethanol
and petroleum leaf extracts of Tinospora cordifolia in
Oreochromis mossambicus showed that all the dose of ethanol
extract significantly enhanced neutrophil activity and fishes
injected with petroleum ether or ethanol extract at a dose of 8
mg/kg were found protected against experimental infection
with virulent Aeromonas hydrophila which shows
immunostimulatory nature of Tinospora cordifolia10. Some
findings also suggested that the Tilapia mossambicus fed with
dietary Amla (Phyllanthus emblica) showed TEC, Hb, TLC,
lymphocyte, neutrophils and monocyte counts increased
significantly in D3 diet (2 % ethanolic extract of P. emblica) fed
fishes. Highly significant increase was observed in serum, total
protein and albumin levels in fishes fed with D2 diet (1% ethanolic
extract of P. emblica) and increase in globulin level were
significant11. Some researchers reported immunostimulatory
potential of Plumbago rosea in Catla catla, post challenged
with Aeromonas hydrophila. For 14 days and they observed an
increase in TEC (P<0.05) and TLC (P<0.05) in the
immunostimulant administered (IS) diet and also Increase in
lymphocyte count12.
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