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International Journal of Clinical Case Reports 2013, Vol.3, No.6, 31
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drugs but such drug resistant isolates were found very
sensitive to sponge extracts. From the present study it
is concluded that the 16
th
and 18
th
fractions of the
sponges
Aurora globostellata
and
Spirastrella inconst-
ans
var.
moeandrina Dendy
respectively has rich
bioactive compounds and that c ompound must be
identified exactly. This will be a panacea for many
pandemic problems.
4
Material and methods
4.1
Isolation of bacteria from catheters
Urinary catheters from patients being cared for in
hospitals in Chidambaram, India w ere collected.
Sections of 1~2 cm were cut from the tip of catheter
that was removed from patients in whom the catheter
was implanted for over 2 months. Forty catheters were
used. The tips were suspended in quarter-strength
Ringer’s solution in (10 mL) in sterile universal
containers, as explained by Sonification for 5 min at
35
kHz in a transonic water bath and by vortex mixing
for 2 minutes was used to remove and disrupt the
colonizing biofilms. The result ing cell suspensions
were cultured onto Cystine lactose electrolyte defici-
ent media (CLED) Chromogenic UTI and tryptone
soya agars (Hi-media). After 24 h incubation the
resulting colonies were identified using standard
procedures. From the catherters seven types of were
isolated viz.,
Proteus mirabilis
,
Escherichia coli
,
Staph
ylococcus epidermis
,
S. aureus
,
Pseudomonas aerugi-
nosa, Neisseria gonnorhaea
and
Candida albicans
.
4.2
Anti-bacterial assay
Disc diffusion method was adopted to screen the
anti-bacterial potential of the extract of sponge against
biofilm isolates on L uria Bertani (LB) agar pla tes.
This is a valuable and inexpensive test to demonstrate
the susceptibility of a particular compound against
pathogens by measuring the relative zone of inhibition.
To achieve this, sterile antibiotic disks (HIMEDIA
Laboratories, India) of 6 mm diameter were loaded
with 20 µL, 40 µL, 60 µL of the extract of the sponge
A. globostellata keeping a control using Tetracycline.
The disks were then air-dried aseptically. Exponential
bacterial cultures were sw abbed on to the LB agar
plates and the disks were left on the agar plates. The
plates were incubated at 37°C for 24 h, meanwhile the
experiments were performed in triplicates.
Acknowledgement
Authors are thankful to Dr. A.J.A.Ranjit Singh, Principal, Sri
Paramakalyani College, Alwarkurichi, Tirunelveli, Tamilnadu,
lab facilities and encouragement. We thankful for supporting
agency of Department of Science and Technology, Govt. of
India, New Delhi.
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