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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, RVS Coimbatore - 641 402,
Tamil Nadu, India *Corresponding author:
balashivam@gmail.com Issued Abstract 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). Introduction 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
population. In 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. Plant Description 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
throughout MATERIALS
Plant
collection Fresh
plant leaves of S. campanulata and T.procumbens
were collected randomly from the gardens and villages of Plant
extraction 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 Bacterial strains used in this study were the
isolated pathogens isolated from clinical cases of bovine mastitis such as coagulase positive Staphylococcus
aureus, coagulase
negative Staphylococcus aureus ( Antibacterial
activity 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 μl 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 μl 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 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. References 1. Ali,M., Rawinder,E.,
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as retrieved on 30. Yadawa, R.N., Saurabh, K., 1998. A new flavones glycoside: 5,7,4 –Trihydraxy – 6, 3 – dimethasey Falavone 5-0 alpha –L- rhamnopyramoside from the leaves of Tridax procumbens Liun. J. Asian Nat. Prod. Res. 1(2), 147-52. Table
1
Antibacterial activity of methanolic and aqueous
extracts of Spathodea campanulata and Tridax procumbens
against bovine mastitis pathogens.
NA-No
Activity Table 2 Phytochemical
screening of Spathodea campanulata and Tridax procumbens.
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