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Effect of Chlorine on the development of marine
biofilms dominated by diatoms
Under the Guidance
of
Dr.Jagadish.s.Patil
Marine Corrosion & Materials Research Division
National institute of oceanography
Goa, India
Jagadeesan v
08MBT03
Synopsis
• Introduction
• Materials and methods
• Results
• Summary
• References
Biofouling- An Intro
•
Biofouling is serious problem of rapid
colonization of aquatic species on immersed
surfaces in an aquatic environment.
•
It is a serious problem in ship hulls, power plant
cooling systems, aquaculture systems, fishing
nets, pipelines, submerged structures and also
oceanographic research instrumentation.
•
Many of the technologies implemented to solve
the current problem and he results are under
the investigation., and remain unsolved.
•
In which chlorination is one of the potential
method of controlling the biofouling both by
industrial and environmental applications.
•
Chlorine is biocide and it is a disinfectant used
to reduce the prevalence of pathogens in
drinking water by municipal water supply.
Aims & objectives
• To prevent biofouling using chlorine as an antifouling agent.
• To evaluate the efficiency of chlorine against mono algal and multi
algal species.
• To find the effectiveness of chlorine against the chlorophyll auto
fluorescence, Biofilm EPS and etc.,
• The evaluation of pulse chlorination on the development of
biofouling.
Diatoms are early colonizers
• Diatoms are the earliest eukaryotic colonizers of submerged surfaces and
one of the most conspicuous organisms in biofilms. The diatoms attach to
the substratum by secretion of EPS (Eg. Wetherbee et al. 1998), and
contribute significantly to the biofilm (Patil and Anil 2005).
• In the present investigation the important biofouling diatoms Amphora and
Navicula (Wetherbee 2008) were evaluated their biofouling potential against
the chlorination along the mixed natural species.
• They were subjected to grow under laboratory with the help of cover glass
substratum, and they start to colonize the substrate at a immediate effect
and they are one of the initial colonizers in the complex mechanism of
biofouling .
Images of mono & multi species biofilm
Natural biofilm
Amphora biofilm
Navicula biofilm
Materials and Methods
• Effect of chlorination on the growth of biofilm diatoms
• Effect of chlorine on chlorophyll fluorescence
• Effect of chlorine on exo polysaccharide (EPS)
• Effect of chlorine on cell structure of fouling diatoms
• Pulse chlorination
• Effect of pulse chlorination on chlorophyll, biofilm development
• Effect of chlorination in the optical transmission
Results
Cell counting Vs chlorination
EPS ASSAY- Alcian Blue
method
Effect of chlorine on the Chlorophyll- Epifluorecscence
analysis
Chlorophyll fluorescence - Navicula
Effect of chlorine on the chlorophyll
•
Effect of chlorine on the chlorophyll is
once again reflected here in the
chlorophyll estimation.
•
The biofilm cover glass are sampled and
chlorophyll analysis was achieved by
acetone 90% extraction method
•
The experiments were done by both
control and with the treatment.
•
The control group is a biofilm without the
treatment of chlorine to compare the
reduction obtained by the chlorine treated
biofilm samples
Live Amphora control
Amphora chlorine treated 0.5%
0.5% 1 min exposure
0.5% 5 min exposure
Cl 1%
1 % 1 min exposure
1 % 5 min exposure
Navicula control
Cl 0.5%
0.5% 1 min exposure
0.5% 5 min exposure
Cl 1%
1 % 1 min exposure
1 % 5 min exposure
Cell counting Vs chlorination Vs algal seedling
Species richness in Natural biofilm Vs chlorination
Pulse chlorination
• Pulse chlorination is a technique that is used to chlorinate the
biofouling candidates at a short interval of time to remove them and
effectively prevent them colonizing.
• In this study chlorine treatment was given in pulse (i.e. at every 6
hours) in order to derive optimal anti fouling results with minimal
chlorine amounts.
• The intermittent chlorination results in the continuous removal
biofouling candidates in a short successive chlorination ensuring the
removal adhered algal species on the surfaces.
Effect of pulse chlorination Biofilm biomass & chlorophyll
Effect of pulse chlorination in optical transmission
• Optical transmission is an
important property in the marine
biosensors.
• The fixed marine biosensors is
often fouled by the activity of
algal species, which is a serious
problem in monitoring the marine
environment.
• The pulse chlorination was
achieved with the clean glass
slides for the transmission
analysis, the transmission was
estimated using the image pro
software in microscope.
Summary
• Chlorination is one of the important antifouling strategy and it is
implemented easily on industrial and environmental application.
• The application of chlorination leads to the death of the algal cells
reflected in the scanning electron microscope analysis.
• which is subsequently confirmed by the cell counting experiment.
• The chlorophyll reduction and Eps analysis in electron microscopy
analysis reveals that it is a most promising agent.
• The pulse chlorination is further earning credit by application of on
the local biosensors applications, to reduce their impact on the
chlorine.
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