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Comparative efficacy of anti-microbial activity of Tinospora cordifolia,

Azadirachta indica and Andrographis paniculata extracts against gram positive
and gram negative bacteria

Article  in  The Indian veterinary journal · May 2017

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 G. Singh et al.
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progesterone assays to monitor pregnancy and determine the
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Mann, G.E. and Lamming, G.E. (2001) Relationship between
Berber, R.C., Madureira, A.E.H. and Baruselli, P.S. (2002)
maternal endocrine environment, early embryo development
Comparison of two Ovsynch protocols (GnRH) for xed timed
and inhibition of the luteolyticmechanisms in cows. Anim.
insemination in buffalo (Bubalus bubalis). Theriogenology, 57:
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Neglia, G., Gasparrini, B., Di Palo, R., De Rosa, C., Zicarelli,
Campanile, G., Di Palo, R., Neglia, G., Vecchio, D. and Gas-
L. and Campanile, G. (2003) Comparison of pregnancy rates
parrini, B. (2007) Corpus luteum function and embryonic mor-
with two oestrus synchronization protocols in Italian Mediter-
tality in buffaloes treated with a GnRH agonist, hCG and pro-
ranean Buffalo cows. Theriogenology,60 (1): 125-33.
gesterone. Theriogenology,67: 1393-1398.
Paul, V. and Prakash, B.S. (2005) Efcacy of the Ovsynch
Francesco, S.Di., Neglia, G., Vecchio, D., Rossi, P., Russo,
protocols for synchronization of ovulation and xed time arti-
M., Zicarelli, L., Occhio, M.J.D. and Campanile, G. (2012)
cial insemination in Murrah buffaloes (Bubalus bubalis). The-
Inuence of season on corpus luteam structure and function
riogenology,64: 1049–1060.
and AI outcome in the Italian Mediterranean buffaloes (Bubal-
us babulis). Theriogenology,78: 1839-1845. Vecchio, D.,Neglia, G., Di Palo, R., Prandi, A., Gasparrini, B.
and Balestrieri, A. (2010) Is a delayed treatment with GnRH,
Ghuman, S.P.S., Dadarwal, D., Honparkhe, M., Singh, J.
hCG or progesterone benecial for reducing embryonic mor-
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against progesterone of application in radioimmunoassay.
Ind. Vety. J., 86: 909-911.

Indian Vet. J., May 2017, 94 (05) : 25 - 28

Comparative Efcacy of Anti-microbial Activity of Tinospora cordifolia,

Azadirachta indica and Andrographis paniculata Extracts Against
Gram Positive and Gram Negative Bacteria
Shraddha Nety1, K.M. Koley, Mamta Choudhary, M.R. Poyam and Durga Chourasia
Department of Veterinary Pharmacology and Toxicology, College of Veterinary Science and Animal Husbandry,
Chhatisgarh Kamdhenu Vivshwa Vidhyalaya (CGKV), Anjora - 491001, Durg (C.G.).
(Received : 12-08-2016 327/16 Accepted : 22-09-2016)

Abstract (AIE) Adrographis paniculata extracts.

The present study describes the antimicro-
bial activity of Tinospora cordifolia (stem),
Azadirachta indica (leaves) and Andrographis
paniculata (aerial parts). Extracts of plants were
obtained by Soxhlet’s extraction in the mixture
of 50%methanol and 50% water. The antibacte-
rial activities were assessed by measuring the
diameter of the inhibition zonesand MIC values
and reported.
Key words : Antibacterial, activity Tinospora
cordifolia extract (TCE), Azadirachta indica
Corresponding author : Email : dr_shraddha20@yahoo.co.in
1
Plants are used in traditional and
Ayurvedic systems of medicine as well as in
tribal medicine in India. Andrographispanicu-
lata (Kalmegh),Tinosporacordifolia (Giloy) and
Azadirachtaindica (Neem) have therapeutic
value due to their mechanisms of action and
presence of phytochemicals. Tinospora cordi-
foliahas been reported to possessantifungal
and anti- microbialactivitiesetc(Sandhuet al.
2013). Azadirachta indicais reported to
have antimicrobial (Reddy etal., 2013) and
Andrographis paniculata reported to posses-
santibiotic, immunomodulatory (Meenachisun-

The Indian Veterinary Journal (May, 2017) 25

 Comparative Efcacy of Anti-microbial ...

Table I. Zones of inhibition of bacteria shown by different medicinal plant extracts and ciprooxacin.

Extract in Each Zones of Inhibition (mm) ± SE

Name of Extract
Disc( mg/ml) Staph. aureus S. gallinarum P.aeruginosa E .coli
6.8mg/ Disc 12.16 ± 0.61(R) 20.0 ± 0.51 (S) 20.0 ± 0.51(R) 12.16 ± 0.54(R)
Tinospora cordifolia (TCE)
13. 0mg 14 ± 0.57(R) 21.00 ± 0.51(S) 25.00 ± 0.44(R) 14.16 ± 0.60 (R)
6.00mg 12.33± 0.61(R) 16.00 ± 0.25(IS) 16.33± 0.42(R) 10.66 ± 0.42(R)
Azadirachtaindica
11.00 mg 16.0 ± 0.25 (R) 18.0 ± 0.44 (S)) 18.0 ± 0.44(R) 12.0 ± 0.44(R)
7.20mg 15.16 ± 0.30 (R) 22.16 ± 0.16 (S) 16.00 ± 0.25(R) 14.33± mm(R)
Andrographis Paniculata (APE)
13.00mg 16.0 ± 0.25 (R) 23.0 ± 0.44 (S) 24.0 ± 0.44 (IS) 16.0 ± 0.51 (IS)
Ciprooxacin 5mcg / Disc 17.00 ± 0.25 (R) 24.00 ± 0.66 (S) 30.33 ± 0.61 (S) 18.30 ± 0.50 (S)

S= Sensitive; IS=Intermediate sensitive; R= Resistant E.coli and Salmonella gallinarum

Resistant ≤ 15mm; Intermediate -16-20 mm; Sensitive -≥ 21 (CLSI., 2014)Staph. aureus-
Resistant m ≤ 19mm; Intermediate -20-21 mm; Sensitive - ≥ 22.

darametal.,2009) and the study was conducted
to evaluate the same.
Materials and Methods
Pure cultures of pathogenic strains of Esche-
richi coli, Staphylococcus aureus, Salmonella
gallinarum and Pseudomonas aeruginosa were
originally obtained from Chandigarh (IMTch)
were used to assess the antibacterial activity of
different extracts was studied by Disc Diffusion
Technique (Baeur,etal.,2014).Five lter paper
blank discs were measured and weight after
each disc was impregnated with a particular
concentration of an extractto calculatethe
netcontent of extract in each disc for different
concentrations. The blank discs were separately
impregnated with distill water only ( 100mg/
ml concentration only) Hydro-alcoholic extracts
of TCE, AIE, and APE were prepared at three
different concentrations of 100mg/ml, 500 mg/
ml and 1000 mg/ml in distill water and were
used for the preparation of extracts impregnated
discs of each extract. 100mg/ml of TCE, AIE and
APE which resulted in lter paper discs contain-
ing 1.80 mg, 2.60 and 2.8 mg Similarly,500 mg/
ml of TCE, AIE and APE which resulted in
lter paper discs containing 6.80 mg, 6.00 mg
and 7.20 mg of TCE, AIE and APE per dischow-
ever, in 1000 mg/mlsolution of TCE, AIE and
APE resulted in extract impregnated lter

Fig 1. Zones of inhibition of T. cardifolia, A. indica and A.
paniculata plant extract and reference antibiotic ciprooxacin
against S. gallinarum. 1. Blank disc with distill water 2. APE
(2.8 mg/disc 3.TCE 1.8 mg/disc) 4. AIE 2.6 mg/disc.
Fig 2. Zones of inhibition of T. cardifolia, A. indica and A.
paniculata plant extract and reference antibiotic ciprooxacin
against S. gallinarum. 1. APE (13 mg/disc), 2. APE (7.20mg/
disc),3.TCE 6.8 mg/disc), 4. TCE 13mg/disc, 5. AIE 6.0 mg/
disc, 6. AIE 11.0mg/disc, 7. Standard ( Ciprooxacin).

26 The Indian Veterinary Journal (May, 2017)

 Shraddha Nety et al.
Fig 3. Zones of inhibition of T. cardifolia, A. indica and A. pan-
iculata plant extracts and reference antibiotic ciprooxacin
against Staph. Aureus.1. APE (13 mg/disc), 2. AIE 6.0 mg/
disc,3. AIE 11.0 mg/disc 4. TCE 6.8 mg/disc), 5. APE (7.20
mg/disc, 6. TCE 13.0 mg/disc, 7. Standard ( Ciprooxacin)
Fig 5. Zones of inhibition of T. cardifolia, A. indica and A. pan-
iculata plant extracts and reference antibiotic ciprooxacin
against E. coli.1. APE (13 mg/disc), 2. APE (7.20 mg/disc).
3.TCE 6.8 mg/disc), 4. AIE 6.0 mg/disc,5. TCE 13mg/disc,
6. AIE 11.0mg/disc, 7. Standard ( Ciprooxacin).
paper discs containing 13.00mg, 11.00 mg and
13.00 mg of TCE, AIE and APE per disc. Each
of four bacteria was grown in nutrient broth,
incubated at 370C overnight and diluted (10-5)
using sterile nutrient broth. Taken one ml of the
broth culture of each bacterium was spread over
the nutrient agar in petri-dishes aseptically.
The extract impregnated discs and a reference
antibiotic (ciprooxacin) disc were placed on the
inoculated nutrient agar in the petri-dishes and
incubated at 370C for 24 hrs and observed for
the presence of zones of inhibition around the
discs and compared to assess the antibacterial
activity of each extract.
The Indian Veterinary Journal (May, 2017)
Fig 4. Zones of inhibition of T. cardifolia, A. indica and A. pan-
iculata plant extracts and reference antibiotic ciprooxacin
against P. aeruginosa. 1. APE (13 mg/disc), 2. AIE 6.0 mg/
disc,3.TCE 6.8 mg/disc), 4.TCE 13mg/disc,5. AIE 11.0 mg/
disc 6. APE (7.20mg/disc),7. Standard ( Ciprooxacin).
Results and Discussion
The TCE, AIE, APE and Standard drug (Cipro-
oxacin) were screened for antibacterial activity
against pathogenic strains of four bacteria viz.
Staphylococcus aureus, Salmonella gallina-
rum, Pseudomonas aeruginosaand Escheth-
erichia coli by disc diffusion method.100mg/ml
concentration did not show any antibacterial
activity against any of the above bacteria.The
study revealed that the TCE and AIE exhib-
ited antibacterial activity against Salmonella
gallinarum. APEshowedsignicant antibacte-
rial activity against Pseudomonas aeruginosaat
high concentration, Salmonella gallinarum
(atboth high and low concentrations) and E. coli
at high concentration but it did not show any
antibacterial activity against Staphylococcus
aureus even at high concentration. Salmo-
nella gallinarum, Pseudomonas aeruginosaand
Eschetherichia coli showed sensitivity to cipro-
oxacin (5microgram/disc) however, Staphylo-
coccus aureus showed resistanceto ciprooxacin
at this concentration (Table I). The blank
discs did not have antibacterial activity. The
results of present study was agreement with
Nagaprashanthi et al. (2012) who also reported
anti- microbial activity of Tinospora cordifolia-
against Staphylococcus aureus, E.coli, Klebsialla
PneumoniaandPseudomonas.Similarly,Reddy et
al. (loc. cit) reported the antimicrobial activity of
Azadirachta indica leaf, bark and seed extracts
against human pathogenic bacteria Viz., Staph-
27
 Comparative Efcacy of Anti-microbial ...
ylococcus aureus, Enterococcus feacalis, Proteus
mirabilis andPseudomonas aeuroginosa.Mishra
et al. (2009)reportedantibacterial activity of
ethanol extract of Andrographis paniculata
against both Gram-negative and Gram positive
bacteria.
Summary
Results revealed that TCE and AIE had antibac-
terial activity only against Salmonella gallina-
rum. APEshowedsignicant antibacterial activ-
ity against Pseudomonas aeruginosa at high
concentration, Salmonella gallinarumatboth
high and low concentrationsand E. coli at high
concentration but it did not show any antibacte-
rial activity against Staphylococcus aureuseven
at high concentrations. The antibacterial activ-
ity observed with TCE, AIE and APE might be
due to the presence of many potent compounds
such as reducing sugars, avonoids, terpenes,
phenolics and alkaloids etc.
Indian Vet. J., May 2017, 94 (05) : 28 - 30
Comparitive Productive Performance of Brucella Seropositive and Seronegative
Cattle of an Organized Dairy Herd
Marykutty Thomas1 and Jobin Thomas
Livestock Research Station, Kerala Veterinary and Animal Sciences University, Thiruvazhamkunnu, Palakkad, Kerala, India- 678 601
(Received : 12-10-2015 468/15
Abstract
An investigation was conducted to analyse
the comparative productive performance of
brucella sero-positive and seo-negative cattle
of an organized crossbred dairy herd. Overall
lactation yield, 305-day milk production and
lactation length were signicantly (p≤0.05)
lower among sero-positive cows compared to
seo-negative cattle. These differences are more
marked (p≤0.05) among rst calvers where the
lactation yield of sero positive animals was
lower by 33. 01%. The average daily milk yield
and peak yield were similar in both groups.
1
Corresponding author : Email : marykutty@kvasu.ac.in
28 The Indian Veterinary Journal (May, 2017)
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Shanthi,V. and Nelson,R. (2013) International J. Curr. Micro-
biology and applied science.,ISSN: 2319-7706 2 (6): 190-194.
Accepted : 29-02-2016)
Key words- Brucellosis, dairy cattle, milk
production
Brucellosis in cattle is characterized by
impaired infertility, late term abortion or birth
of weak or dead calves (Olsen and Johnson 2011).
Analysing the impact of brucellosis on production
performance of dairy cattle in organized herds
is imperative to make a conclusive decision on
disease control with limited resources and such
reports are scanty. The present investigation
was intended to assess the comparative produc-
tion performance among brucella sero-positive
and sero-negative cattle in an organized dairy
herd.