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Plan of Work
Conclusion.
What is a biofilm?
A biofilm is any group of microorganisms in which
cells stick to each other on a surface.
Eg: Plaques that form on teeth causing tooth decay is
a type of bacterial biofilm, rocks are coated with
biofilm in a stream or river, gunk that clogs drains,
etc.
Biofilm forms when bacteria adhere to surfaces in
moist environments by excreting a slimy, glue-like
substance.
Over 500 bacterial species have been identified in typical dental plaque
biofilms.
History of Biofilms
The group of Dr. Costerton, in 1978, used the name biofilms as a more
generic term for microorganisms adhering to wet surfaces in freshwater
ecosystems.
Aniridescentbiofilmonthesurfaceofafishtank
Source:
https://en.wikipedia.org/wiki/Biofilm#/media/File:Iridescent_biofilm_on_a_fishtank.JPG
Source:
http://www.biofilm.montana.edu/node/2390
They play key roles in the development of our immune systems and in
the anatomy of the mucosal surfaces, and also provide protective
functions against exogenous pathogens.
Habitat
Biofilms can grow in the most extreme environments: from, for example, the
extremely hot, briny waters ofhot springsranging from very acidic to very
alkaline, to frozenglaciers.
In the domestic environment, biofilms can grow inshowers. Biofilms can form
inside water and sewagepipesand cause clogging and corrosion.
Biofilms are found on the surface of and inside plants. They can either
contribute to crop disease or, as in the case of nitrogen-fixing Rhizobium on
roots, exist symbiotically with the plant.Examples of crop diseases related to
biofilms include Citrus Canker, Pierce's Diseaseof grapes, and Bacterial Spot
of plants such as peppers and tomatoes.
Source:
https://en.wikipedia.org/wiki/Biofilm#/media/File:Iridescent_biofilm_on_a_fishtank.JPG
Dental plaque
Dental Plaque
Source: http://www.redrockdental.org/lp/plaque.html
Legionellosis
Source: http://biomedfrontiers.org/infection-2013-dec-2/
Source: http://m2002.tripod.com/legionellosis.htm
ANTIMICROBIAL
RESISTANCE
INTRINSIC
The biofilm matrix
may act as a
diffusion barrier
Establishment of
microenvironment
s within biofilms
Differentiation into
persister cells
Increased
production of
oxidative stress
EXTRINSIC
Biofilms can act as physical diffusion barriers to prevent antibiotics from reaching their
targets. Antibiotic was shown to be able to penetrate these structures through a thick
mixture of exopolysaccharide, DNA, and protein in order to reach the targets.
However, this mixture was not able to achieve an effective concentration in some all parts
due to the physical and/or chemical properties of the matrix, which resulted in an apparent
increase in resistance.
Recent mathematical modeling has predicted that while limited antibiotic diffusion may lead
to the death of the outer layer of bacteria, it also stimulates subpopulations of bacteria
buried deeper within the biofilm to enact adaptive changes, thereby countering the insult.
However, limited antibiotic diffusion does not appear to be a universal trait shared by all
biofilms, and, there is conflicting data about whether the biofilm matrix is a major
contributing factor influencing biofilm resistance.
The depletion of nutrients and oxygen inside biofilms might cause an altered metabolic
activity and lead to slow growth of the bacteria. Several studies have revealed oxygen
limitation and the presence of hypoxic zones deep within biofilms, with nutrient diffusion
through biofilms being restricted.
Inspection of environmental as well as in vitro biofilms has revealed that the oxygen
concentration may be high at the surface, but low in the centre of the biofilm where
anaerobic conditions may be present.
Likewise, growth, protein synthesis and metabolic activity are stratified in biofilms, for
example, there is a high level of activity at the surface but a low level in the centre, resulting
in slow or no growth. This fact is one of the explanations put forward for the reduced
susceptibility of biofilms to antibiotics.
In general, all antimicrobials are more effective in killing rapidly growing cells. For example,
penicillin and ampicillin have an absolute requirement for cell growth to occur in order to kill,
be able to, with the mortality rate being proportional to the rate of growth.
Persister cells are considered to be either nongrowing or slow-growing, and also have a
greatly reduced susceptibility to antibiotics.
In the persisters theory, these small subpopulations of bacteria within the biofilms
differentiate into dormant cells, are able to survive extreme antibiotic treatment and have
been hypothesized to be the result of phenotypic variations rather than due to stable genetic
changes.
The formation of persisters might represent a common mechanism used by a wide range of
bacteria during biofilm formation. This persistent population within biofilms may drastically
inhibit the complete eradication of the biofilms, even after prolonged, high-level antibiotic
treatment.
However, it is unclear what relationship exists between planktonic persisters and biofilm
resistance, and the mechanisms(s) by which persisters form and/or confer increased
antibiotic tolerance are still unknown.
Oxidative stress is caused by an imbalance between the production of oxidants, such as the
free radicals, peroxide and nitric oxide, with the levels of antioxidant defenses.
Oxidative stress is considered to cause enhanced mutability in biofilms. Recent data suggest
that microcolonies structures, due to endogenous oxidative stress, are specific sites within
biofilms where enhanced genetic adaptation and evolutionary change take place.
In addition, Boles and Singh showed that endogenous oxidative stress in biofilms promotes
antibiotic resistance and that the addition of antioxidants reduces the diversity in biofilms.
The source of -lactamase in biofilms has been considered to come from the
layer of lysed bacteria resulting from exposure to antibiotic, and is related to
the release of defensive enzymes into the extracellular space.
ANTIBIOTIC
MICROORGANISM
PENETRATIO REFERENCE
N?
Piperacillin
P.aeruginosa
No
Rifampin
S.epidermis
Yes
Dunne et al.,
1993
Vancomycin
S.epidermis
Yes
Gentamicin
P.aeruginosa
No
Yasuda et al.,
1993
Latamoxef
E.coli
Yes
Jouenne et al.,
1994
Ciprofloxacin
P.aeruginosa
Yes
Amikacin
P.aeruginosa
No
Shigeta et al.,
1997
Prevention of biofilm formation may be accomplished by avoiding conditions that lead to cell
attachment and selecting conditions that make the environment unfavorable for microbial
growth.
Regularly-scheduled cleaning and keeping surfaces free of moisture and residue (bacteria
food) is the best defense against biofilms. Once established, biofilms are difficult to get rid of.
Even if the surface looks clean, biofilms can cling tenaciously to out of the way areas.
Studies show that due to their protective matrix, spraying with bleach or another type of
antimicrobial, or a shower or toilet bowl cleaner, and then just rinsing, isnt going to remove
them. Antimicrobial chemicals cannot penetrate the biofilm to kill all bacteria.
Like dental plaque, biofilm must be agitated, broken up, and removed before an antimicrobial
chemical (in the example of plaque - mouthwash) will be effective.
Biofilm removal and inactivation is achieved by combining proper cleaning and sanitizing
agents, adequate exposure time, proper temperature and mechanical action.
This combination dissolves the biofilm and the organic material to which it adheres, allowing
the sanitizer to inactivate the released, sensitive cells. Extensive scrubbing with proper
chemicals is important in biofilm removal.
Coating agent
Coating method
Mechanism
Antibiotics
Bactericidal/Bacteriostatic
Silver
Bactericidal
Furanones
Bactericidal/Bacteriostatic
QAS
Covalent bonding
Covalent bonding
Bactericidal/Bacteriostatic
TMS
Anti-adhesion
PLL-g-PEG
Anti-adhesion
pCBMA
Anti-adhesion
Anti-adhesion
Anti-adhesion
Selenocyanatodiacetic acid
Covalent bonding
Anti-adhesion
Surface grafting
Anti-adhesion
Conclusion
Discovery of these factors should lead to new and better treatments for
biofilm related infections. New strategies are required to overcome
extreme biofilm antibiotic resistance through the development of novel
therapies aimed at disrupting biofilms and killing the constituent
bacteria, with manipulation of intrinsic and extrinsic resistance
pathways providing much promise for future treatment of biofilm
infections.
Proper cleaning and sanitation work best for biofilm prevention while its
removal is only necessary if prevention fails.
References
Costerton JW, Geesey GG, Cheng GK. How bacteria stick. Sci. Am. 1978; 238:8695
Donald RM. Biofilms: Microbial life on surfaces. Emerg Infect Dis. 2002; 8(9): 881890.
Costerton JW, Lewandowski Z, Caldwell DE, Korber DR, Lappin-Scott HM. Microbial
biofilms. Annu Rev Microbiol. 1995; 49:711745.
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