ABSTRACT   Bovine Mastitis is an intramammary infection which is most common among the dairy cattle and continues to be the most costly disease to the dairy farmers. Presently, antibiotics are used for treatment of mastitis leading to the development of antibiotic resistant strains and consumer health problem. The present study investigates in vitro antibacterial activity of two medicinal plants namely Cymbopogon citratus and Punica granatum L against causative organisms of bovine mastitis. Aqueous and methanol extracts of two plants were investigated by agar well-diffusion method. Methanol and aqueous extracts of Cymbopogon citrates and Punica granatum L.  were found to possess potency against infectious pathogens Staphylococcus aureus, Coagulase negative Staphylococcus aureus, Streptococcus uberis and Escherichia coli isolated locally from clinical and sub clinical samples of Mastitis from Karnataka region whereas .Phytochemical screening of the plants revealed mainly the presence of alkaloids, tannin, saponin, terpenoids and falvonoids. Keywords: Cymbopogon citratus, Punica granatum L, Aqueous extracts, methanol extracts, phytochemical screening, antibacterial activit

Deshpande, Mruganka

nath, gopi

T. B, Suneetha

English

Advanced Research Journals

International Journal of Phytomedicine 3 (2011) 333-337 
http://www.arjournals.org/index.php/ijpm/index 
 
Original Research Article 
              
               
Preliminary Analysis of Two Medicinal Plants against causative organism 
of Bovine Mastitis 
Gopinath S.M1*, Suneetha T.B1, Mruganka1, V.D, Ananda S1, 
 
*Corresponding author: 
 
Gopinath.S.M 
1. Department of 
Biotechnology, Acharya 
Institute of Technology, 
Bangalore- 90 
Email:gopinath@acharya.ac 
 
 
 
 
 
 
 
 
 
 
 
 
Abstract 
Bovine Mastitis is an intramammary infection which is most common 
among the dairy cattle and continues to be the most costly disease to the 
dairy farmers. Presently, antibiotics are used for treatment of mastitis 
leading to the development of antibiotic resistant strains and consumer 
health problem. The present study investigates in vitro antibacterial 
activity of leaves of two medicinal plants namely Cymbopogon citratus 
and Punica granatum L against causative organisms of bovine mastitis. 
Aqueous and methanol extracts of two plants were investigated by agar 
well-diffusion method. Methanol and aqueous extracts of leaves of 
Cymbopogon citrates and Punica granatum L.  were found to possess 
potency against infectious pathogens Staphylococcus aureus, Coagulase 
negative Staphylococcus aureus, Streptococcus uberis and Escherichia 
coli isolated locally from clinical and sub clinical samples of Mastitis 
from Karnataka region whereas .Phytochemical screening of the plants 
revealed mainly the presence of alkaloids, tannin, saponin, terpenoids 
and falvonoids.  
Keywords: Cymbopogon citratus, Punica granatum L, Aqueous 
extracts, methanol extracts, phytochemical screening, antibacterial 
activity 
Introduction 
Bovine Mastitis is an intramammary infection 
which is most common among the dairy cattle. 
This disease is seem to reduce milk yield,  
producers profits and milk product quality. 
Microbiological causes of mastitis are many and 
multiple factors involved in the management, 
housing, milking should be considered and 
continues to be the costliest disease in the dairy 
industry all over the world [1].The repeated use 
of antibiotics to treat Mastitis for a long period 
may cause multidrug resistivity in causative 
organisms which requires high doses of 
antibiotics may leads to accumulation of large 
amount of antibiotics in milk and its products, 
again a potential hazard [2]. The World Health 
Organization (WHO) noted that the majority of 
the world's population depends on traditional 
medicine for primary healthcare. Medicinal and 
aromatic plants are widely used as medicine and 
constitute a major source of natural organic 
compounds. [17] Some medicinal plants have 
been used for a wide variety of purposes such as 
food preservation, pharmaceutical, alternative 
medicine, and natural therapies for many 
thousands of years. It is generally considered that 
compounds produced naturally, rather than 
synthetically, will be biodegraded more easily 
ISSN: 0975-0185 
Gopinath et al. International Journal of Phytomedicine 3 (2011) 333-337 
 
334 
 
and therefore be more environmentally 
acceptable. Positive response of plant based drugs 
might lies in the structure of natural products 
which react with toxins and / or pathogens in 
such a way that less harm is then to other 
important molecules or physiology of the host. It 
is because of this reason drug designing studies 
now-a-days having come up as a new field of 
research [3].  
 
In regard to above information’s, the present 
study was undertaken to investigate the effects of 
aqueous and methanolic leaf extracts of 
Cymbopogon citrates and Punica granatum L.  
This study is to compare the mechanism of in 
vitro antibacterial action of the two plant 
materials against bovine mastitis isolated 
pathogens. 
 
Plant description 
Punica granatum L. commonly known as 
Pomegranate belongs to the Family Punicaceae. 
The pomegranate is native to the Iranian Plateau, 
the Himalayas in north Pakistan and Northern 
India 
Punica granatum is a shrub or small tree with 
several upright, thorny stems, the leaves are 
elliptic, roughly 2x1 inches. The plant has also 
been used as an antispasmodic and 
antihelmintic.[16] 
 
Cymbopogon citrates, commonly known as 
lemon grass as well as oil grass is a native herb 
from India and is also cultivated in other tropical 
and subtropical countries. It is a tall perennial 
grass. It is consumed as an aromatic drink and 
used in traditional cuisine for its lemon flavor [4].  
 
 
Materials and Methods 
The leaves of both the plants were collected from 
Acharya Institute of Technology campus, 
Soladevanahalli, Bangalore, India in April 2011. 
The dried leaves were grinded into fine powder 
and the total mass was subjected for extraction by 
hot percolation method with water and Methanol 
in soxhlet apparatus for 72 hrs. Each solvent 
extraction step was carried out for 24 hrs and 
after extraction the extracts were concentrated to 
form a thick solution by keeping in hot water bath 
and then stored at 4°C for further study[7]. 
Bacterial strains 
Bacterial strains used in this study were isolated 
from clinical cases of Bovine mastitis, namely 
Staphylococcus aureus, Streptococcus uberis, 
Escherichia coli, and coagulase negative 
Staphylococcus aureus (CONS). All the strains 
were confirmed by cultural and biochemical 
studies [5].These organisms were cultured on 
Nutrient agar at 37 ºC for 24hrs and maintained 
in nutrient agar slants at 4ºC for further use. 
Antibacterial activity  
The antibacterial assay of aqueous and 
methanolic extracts was performed by agar disc 
diffusion method [8]. The molten Mueller Hinton 
agar was inoculated with 100µl of the inoculums 
(1*106 CFU/ml) and poured into the petriplate 
(Himedia). For agar disc diffusion method, the 
disc (0.7cm), (Himedia) was saturated with 100µl 
of the test compound, allowed to dry and was 
introduced on the upper layer of the seeded agar 
plate. The plates were incubated overnight at 
37ºC. Microbial growth was determined by 
measuring the diameter of the zone of inhibition 
of each bacterial strain. 
Phytochemical analysis 
Phytochemical analysis for major 
phytoconstituents of both the plant extracts was 
undertaken using standard qualitative methods as 
described by various authors [9] [10]. The plants 
extracts were screened for the presence of 
biologically active compounds like alkaloids, 
phenolics, tannins, flavonionds, saponins 
phenols, quinone, anthraquinone, steroids etc. 
 
 
Gopinath et al. International Journal of Phytomedicine 3 (2011) 333-337 
 
335 
 
Results and Discussion 
Table 1 represents the antibacterial activity of 
Cymbopogon citrates and Punica Granatum L. In 
Punica Granatum  L and Cymbopogon citrates, 
the most pronounced activity was shown by 
methanol as well as water extracts against 
staphylococcus aureus and minimum activity was 
observed in streptococcus uberis from aqueous 
and methanol extracts of the 2 plants. Figure 1 
shows the values of zone of inhibition versus 
organism which cause bovine mastitis. Table 2 
represnts the phytochemical screening of the 2 
plants. Most of the secondary metabolites were 
identified in the polar extracts. Alkaloids are one 
of the characteristic secondary metabolite in 
leaves of this genus found in aqueous extract. 
Tannins are water soluble polyphenols known as 
tannic acid acts as antimicrobial agents. Presence 
of tannins is to prevent the development of 
microorganism by precipitating microbial 
proteins [11].  
 
 
 
 
 
 
Table 1: Antibacterial activity of Cymbopogon citratus and 
Punica Granatum L. 
 
 
Organism 
Zone of inhibition (mm) 
Cymbopogon 
citrates (Leaves) 
Punica Granatum 
L. ( Leaves) 
M W M W 
Staphylococcus aureus 20 16.2 36 36 
Streptococcus uberi 16 13 25 25 
Escherichia coli 12 10 27 32 
CONS 16 18 32 30 
 
 
 
 
 
 
 
 
 
 
Fig 1: ZONE OF INHIBITON (mm) vs Organisms for all extracts obtained 
 
  
  
Gopinath et al. International Journal of Phytomedicine 3 (2011) 333-337 
 
336 
 
Phytotherapuetically, tannin containing plants are 
used to treat non specific diarrhea, inflammations 
of mouth, throat and injured skins [12]. 
Flavonoids are known to be synthesized by plants 
in response to microbial infection. Hence it 
should not be surprising that they have been 
found to be effective as antibacterial substances 
against a wide array of infectious agent [13].  
 
 
Table 2: Phytochemical analysis of Cymbopogon citratus and 
Punica Granatum L. 
 
  
Cymbopogon 
citrates (Leaves) 
Punica Granatum L. 
(Leaves) 
Compounds Methanol Water Methanol Water 
Steroids - - - - 
Terpenoids - + - + 
Alkaloids + + + + 
Flavonoids + + + + 
Coumarins - - - - 
Saponins + + + + 
Tannins + - - - 
Phenols + + + + 
Catechin - + - - 
Anthraquinone - - - - 
quinone + + + + 
 
 
All plant pots synthesize some chemicals by 
themselves to perform their physiological 
activity. Methanolic extracts of Cymbopogon 
citrates revealed the presence of alkaloids, 
flavonoids, saponins and phenols. Along with 
these terpenoids and catechin were present in the 
aqueous extracts of Cymbopogon citrates. 
Alkaloids, flavonoids, saponins and phenols are 
present in methanolic extracts of Punica 
granatum L whereas along with these terpenoids 
are seen in water extracts of Punica granatum L. 
The medicinal value of these secondary 
metabolites is due to the presence of chemical 
substances that produce a definite physiological 
action on the human body. The most important of 
these substances include carbohydrates, phenols, 
phytosterols, sterols, terpenes and volatile oil for 
cell growth, replacement, and body building [14] 
[15] describes today’s traditional medicine as 
undoubtedly the oldest form of medicine and 
probably had evolved simultaneously with the 
evolution of human beings. Results suggest that 
extraction with water is shown to posses many 
bioactive compounds than methanol extraction. 
Thus, Water extraction is a better option for our 
analysis [3, 6].  With the traditional knowledge in 
the background potential plants can be prospected 
to reach bioactive compounds which can be 
further formulated. 
 
Conclusion 
 More bioactive compounds have been seen in 
aqueous extract of leaves suggesting water 
extracts to be more powerful solvent for 
extraction. Leaves of Cymbopogon citrates and 
Punica Granatum L have shown to have same 
effect against the causative organism of Bovine 
mastitis and contain potent bioactive compounds 
which have antibiotic effect. But Punica 
Granatum L, is much more effective as compared 
to Cymbopogon citrates. Hence, these plants 
extracts may be used for extracting drug 
molecules to work against Bovine mastitis. 
   
References 
[1] Adaobi Ezike, Godwin Ebi, Peter Akah, 
Uchechukwu Okeudo. 2011. J. Chem. Pharm. 
Res.,2011, 3(3):676-679 
[2] Annapoorani Chockalingam, Dante S. 
Zarlenga, Douglas D. Bannerman., American 
Journalof Veterinary Research. 68 (11): 2007, 
pp: 1151 –59. 
[3] Bauer, A. W., Kirby, W.M.M., Sherris, J. C., 
Turck, M., 1966. Am. J. Clin. Pathol. 45: 493-
496. 
[4] C.K. Hindumathy., 2011. World Academy of 
Science, Engineering and Technology; 193-
197 
Gopinath et al. International Journal of Phytomedicine 3 (2011) 333-337 
 
337 
 
[5] D. H. Tambekar and S. B. Dahikar., 2011 J. 
Chem. Pharm. Res., 2010, 2(5): 494-501 
[6] Egharevba, Henry Omoregie et al., New York 
Science Journal; 2010;3(12); 91-98 
[7] Gopinath. S. M, Suneetha. T. B, Mruganka, 
V. D, Ananda. S; J. Chem. Pharm. Res., 2011, 
3(5):514-518   
[8] Jamine. R,Daisy. P and Selvekumar.B.N., 
2007. Research Journal of 
Microbiology.2(4):369-374 
[9] Klastrup O. Dairy Federation. 85: 1975, pp: 
49-52. 
[10] Kubmarawa D, Khan ME, Punah AM and 
Hassan 2008. Journal of Medicinal plants 
research 2(12):352-355. 
[11] Parekh, J., Chanda, S., 2007b. Afr. J.Biol. 
Res. 10: 175-181 
[12] Pranay Jain, Gulhina Nafis; Journal of 
Pharmacy Research 2011, 4(1), 128-129. 
[13] Prasad, N.R., Viswanathan.S., Renuka Devi, 
J., Vijayashree Nayak., Sweth,V.C., Archana 
parathasarathy, N and Johana Rajkumar., 
2008. Journal of Medicinal Plants 
Research.2:268- 270. 
[14] Sachin Kumar, Hotam Singh Choudhary, 
Chandrabhan Seniya., 2011. J. Chem. Pharm 
Res., 2011, 3(4):854-860  
[15] Sofowora, A., 1993. Recent trends in 
research into African medicinal plants. 
J.Ethnopharmacol. 38: 209-214. 
[16] Trease, G.S. and Evans, H.C., 1978. 
Textbook of pharmacognosy. 9th edition. 
Bailiar Zindall and Co., London. 
[17] Wynn GS.2001.Herbs in Veterinary 
Medicine. Alternative Veterinary Medicine. 
 
 
 


Preliminary Analysis of Two Medicinal Plants against causative organism of Bovine Mastitis

https://www.arjournals.org/index.php/ijpm/article/download/370/356

Preliminary Analysis of Two Medicinal Plants against causative organism of Bovine Mastitis

Abstract

Similar works

Full text

Available Versions

Advanced Research Journals