Ethnobotanical Leaflets 13: 152-58. 2009.
In Vitro Antibacterial Activity of Two Medicinal Plants
against Bovine Udder Isolated Bacterial Pathogens
from Dairy Herds
M.P. Ayyappa Das2, R. Dhanabalan1* and A. Doss1
1Department of Microbiology
2Department of Biotechnology,
Coimbatore - 641 402, Tamil Nadu, India
*Corresponding author: firstname.lastname@example.org
Bovine mastitis continues to be the most costly disease to the dairy farmers. In Tamil Nadu dominates as one of the most prevalent diseases in dairy cattle among the dairy farms. Mastitis treatment with antibiotics leads to the development of antibiotic resistant strains and consumer health problem. The present study is an in vitro antibacterial activity of two medicinal plants against bovine udder isolated bacterial pathogens. Aqueous and methanol extracts of two plants were investigated by agar disc and well-diffusion method. Methanol extracts of Tridax procumbens and Spathodea campanulata showed significant activity against coagulase positive Staphylococcus aureus (8.0 � 0.70) and Streptococcus agalactiae (7.6 � 0.54) respectively. Phytochemical screening of the plants revealed the presence of alkaloids, tannin, saponin, steroids, terpenoids and falvonoids.
Keywords: Spathodea campanulata; Tridax procumbens; Antibacterial activity; Bovine Mastitis; Ethno-Veterinary Medicine (EVM).
Mastitis continues to be the most costly
disease to the worldwide dairy industry and research efforts continue toward
development of improved methods for the control and elimination of this
disease from dairy herds (Salmon et al.,
1997). The use of antimicrobials over long periods has triggered the development
of multidrug resistant strains, which has resulted
in the use of increasing doses of antimicrobials, causing the danger of
increasing amounts of drug residues in milk, a potential biohazard. Medicinal
and aromatic plants have played an important role in the socio-cultural,
spiritual and healthcare needs of the rural and tribal people and their live
stocks in the emerging and developing countries. In many developing
countries, a large section of population relies on traditional system of
medicines derived from medicinal and aromatic plants to meet not only their
own healthcare needs but for their live stocks also. Traditional medicine has
existed since pre-historic times and flourishes today as the primary form of
human and animal medicine for perhaps as much as 80% of the world's
In view of the dearth of above information�s, the present study was undertaken to investigate the effects of aqueous and methanolic leaf extracts of Spathodea campanulata and Tridax procumbens. This study is to elucidate the mechanism of in vitro antibacterial action of plant material against bovine mastitis isolated pathogens.
Spathodea campanulata P.Beauv is a
species belonging to the Bignoniaceae family,
native from equatorial
�Tridax procumbens Linn (Compositae)
is common grass found in tropical areas of all countries, growing primarily
during raining season. It habitats waste places, road sides and hedges
plant leaves of S. campanulata and T.procumbens
were collected randomly from the gardens and villages of
For aqueous extraction, 10 g of air-dried powder was mixed with 100 ml distilled water and stand at room temperature for 48 h. It was then filtered through 8 layers of muslin cloth and centrifuged at 5000 g for 10 min. The supernatant was collected and stored at 4oC. For solvent extraction, 10 g of air dried powder was mixed with 100 ml of organic solvent (methanol) in a conical flask, plugged with cotton and then kept on a rotary shaker at 190 - 220 rpm for 24 h. After 24 h, it was filtered through 8 layers of muslin cloth and centrifuged at 5000 g for 10 min. The supernatant was collected and the solvent was evaporated using rotary vacuum pump and stored at 4oC in air-tight bottles.
Bacterial strains used in this study were the
isolated pathogens isolated from clinical cases of bovine mastitis such as coagulase positive Staphylococcus
negative Staphylococcus aureus (
The antibacterial assay of aqueous and methanolic extracts was performed by two methods. The agar disc diffusion method (Bauer et al., 1966; Parekh and Chanda, 2006) and agar well diffusion method (Perez et al., 1990; Nair and Chanda, 2005). The Mueller Hinton Agar media, along with the inoculum (108 cfu/ml) was poured into the petri plate. For the agar disc diffusion method, the disc (0.7 cm) (Hi-Media) was saturated with 100 l of the test compound, allowed to dry and then placed on the upper layer of the seeded agar plate. For the agar well diffusion method, a well was prepared in the plates with a cup-borer (0.85 cm) and 100 l of the test compound was pipetted directly into the well. The plates were incubated overnight at 37�C. Antibacterial activity was determined by measuring the diameter of the zone of inhibition (mm) surrounding bacterial growth. For each bacterial strain, controls were included that comprised pure solvents instead of the extract (Parekh and Chanda, 2007b). The experiments were repeated three times and the mean values are presented with � Standard Deviation (SD).
Phytochemical screening was carried out on both the plant extract to identify the���� phyto-constituents (Trease and Evans, 1989).
Results and Discussion
����������� The traditional ethno-veterinary
medicinal practices are being followed by the ruralities
through which a number of veterinary diseases are managed in the developing
countries. The use of antibiotics and other chemical products are banned for
animal healthcare in a number of countries because of human healthcare. The
World Health Organization (WHO) states that 74% of the plants derived
medicines have a modern indication that correlates with their traditional,
cultural (and sometimes ancient) uses (Wynn, 2001). The results of
antibacterial activity of S.campanulata and T.procumbens� shown in table 1. Methanol extracts of S.campanulata
showed significant activity against Streptococcus
agalactiae (7.6�0.547) followed by Escherichia coli (7.2�0.836), Streptococcus uberis
(7.2�0.447), coagulase positive Staphylococcus aureus (7.0�1.0). Whereas only a moderate activity was observed against Klebsiella
pneumonia (5.8�0.447) and coagulase negative Staphylococcus aureus
The phytochemical screening revealed the presence of alkaloids, tannin, saponin, steroids, terpenoids and falvonoids (Table 2). 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 in vitro effective antibacterial substances against a wide array of infectious agents������� (Jamine et al., 2007). Tannins have been reported to prevent the development of microorganisms by precipitating microbial protein and have been reported to have various physiological effects like anti-irritant, antisecretolytic, antiphlogistic and antiparasitic effects���������������������������� (Naveen Prasad et al., 2008). Wynn (2001) describes the today's traditional medicine, as undoubtedly the oldest form of medicine and probably evolved simultaneously with the evolution of human beings. EVM has been a mainstay of developing countries that lack access to conventional medicines for veterinary health care, often only unaffordable means to poor farmers.
The EVM practices could be an effective approach for tackling problems like mastitis, bovine viral diarrhea and many deficiency disorders. With the traditional knowledge in the background potential plants can be prospected to reach the active fraction or molecule(s), which can be further formulated, also the dried plant material itself could be utilized by premixing it with the fodder of cattle feed while utilizing the pure molecule as a marker to maintain the product quality control. Further studies may be necessary to elucidate the specific phytoactive compounds in the leaf extract of the plant S.campanulata� and T.procumbens.
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Table 1 Antibacterial activity of methanolic and aqueous extracts of Spathodea campanulata�
�and Tridax procumbens against bovine mastitis pathogens.
Table 2 Phytochemical screening of Spathodea campanulata� and Tridax procumbens.