Strategies to reduce biofilm formation

IIS(Deemed to be University)
JAIPUR

Submitted by Supervised by:

Ms. Kalpana Dr. Charu Sharma
M.Sc Biotechnology Senior Assistant Professor
Semester IV Department of Biotechnology
Biofilm : Composition and structure
• Biofilms is a thick layer of prokaryotic organism that have aggregated to form a
colony. The colony attaches to a surface with a slime layer (EPS) which aids in
protecting the microorganisms.
• The term ‘biofilm’ was formally introduced in 1978 by Costerton.
• Biofilm is made up of various components (Table1) and all components are
arranged in form of a 3-D like structure ( Rabin et al ., 2015).

Table 1 Composition of biofilm (Jamal et al., 2015)

Components Percentage of matrix
Water 97%
Microbial cells 2-5%
DNA &RNA (only 1 present) <1%

Polysaccharides 1-2%
Proteins <1-2%
Biofilm Formation
• It is a multi step process
• It involves – Attachment, Formation of 3- D structure and maturation and
dispersal

Fig. 1 The biofilm life cycle

Source -
https://www.cs.montana.edu/webworks/projects/stevesbook/contents/chapters/chapter001/sectio
n002/black/page001.html
 • Trickling filters with engineered
biofilm
Waste water • Engineered biofilms involves
Treatment prokaryotic and eukaryotic organisms
as well as higher life forms(Scholz,
2006 )

Positive
roles of
biofilms

• used in cleaning up oil and gasoline

spills
Bioremediation
• A.borkumensis, Alcanivorax ,
Methylococcae( Biello, 2015)
 Negative Roles of Biofilms
(Kokare et al., 2009)

Cystic Fibrosis Otitis media Tooth related

disease
• Formation of thick and • Inflammation occurs in • Dental plaque- Acid is a
sticky mucus, which mucoperiosteal lining major cause of tooth
blocks airway decay
• Peridontitis - Leads to
exfoliation of teeth

• Pseudomonas • S. pneumonia, H. • Streptococcus sanguis,

aeroginosa, S. aureus, influenza, S. epidermidis, Streptococcus gordonii,
H. influenza and P. aeruginosa Streptococcus oralis,
(Kokare et al., 2009). Actinomyces species
• Organism responsible for
the chronic periodontal
is Porphyromonas
gingivalis.
Infections on medical implants :Biofilms are able to colonize on medical devices such as
catheters and implants and cause infections which directly effect on humans health.

Urinary infections - The organisms commonly contaminating these types of

catheters and developing biofilms are S. epidermis, Enterococcus faecalis, E.
coli, Proteus mirabilis, P. aeroginosa, K. pneumonia
(Kokare et al., 2009)

(Source-https://www.dreamstime.com/stock-photos-biofilm-formation-catheter-image25261633)

Prosthetic joint infection - Basically involves hip and knee replacement.

Bacteria causing prosthetic joint infection are usually caused by
Staphylococcus aureus, E. coli, Staphylococci, Priopionibacterium acne
( Song et al., 2013)
(Source-https://medicalxpress.com/news/2017-07-antibiotic-releasing-polymer-eradicate-joint-implant.html)

Prosthetic heart valve infection - Results in the tissue damage, leading

to the accumulation of the platelets. The primarily microorganisms
responsible for this condition are S. epidermis, S. aureus. Streptococcus
spp, Gram negative Bacilli, Enterococci and Candida spp. (Kokare et al.,
2009)

(Source- https://airfreshener.club/quotes/infections-biofilm-humans.html)
Biofilm formation in food industry: Presence of bacterial aggregation leads to unhygienic

conditions that cause economic losses due to the reduced shelf life of the affected food

products, resulting in food spoilage (Bridier et al., 2015) .

• The Staphylococcus biofilm forms on raw meat and on ready-to-eat food product surfaces

(sandwiches, cheeses, puddings, pastries, and so on) release enterotoxins, which cause

severe food poisoning in humans (Kadariya et al., 2014).

• Microorganisms like L. monocytogenes, Yersinia enterocolitica, C. jejuni, Salmonella

spp., Staphylococcus spp., B. cereus, and E. coli are responsible for biofilm formation in

food industry (Anand et al., 2014).

.
Prevention and control strategies
Possible antibiofilm strategies can be based on - inhibition of microbial cells to the
surface, interference with signal molecules and dispersal of matrix. Different
methods (Figure2) are used to prevent the biofilm formation.

Fig. 2The strategies for prevention and treatment of biofilms.

(Source: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6312881/)
• Inhibition of microbial adhesion to the surface and of colonization
 Microbial surfaces are hydrophobic in nature (Van der Mei et al., 1988)
 Adhesion of microbial cells can be prevented by coating the surface with hydrophilic
polymers (Fig. 3 a) and with antibiotic coating (Fig. 3b).

Fig: 3 Surface with hydrophilic coating (a) device surfaces with antibiotic coating (b)
(Van der Mei et al., 1988)
Coatings of hydrophilic Antibacterial coating Electric methods Use of nanoparticles
polymers

Heparin coatings Gentamicin, rifampicin, When DC voltage, low Silver has antibacterial
prevents thrombosis and vancomycin and AC current ,pulsed activity and nanosilver
reduces microbial tobramycin (Van de electric methods are coatings are widely
colonization. Belt et al., 2001). used in combination with applied to medical
antibiotic create devices such as catheters,
synergistic method heart valves and wound
dressings.

 Hyaluronic acid  A continuous  In a study , an electric Gold, diamond, zinc

(Cassinelli et al., delivery of field of (3-4V/cm) oxide and titanium,
2000) used to coat norfloxacin was given given along with coating (Veerachamy et
polyurethane to sessile cells. chloramphenicol al., 2014)
catheters  Growth of Proteus treatment showed a
 poly-N- vulgaris was stopped synergistic effect and
vinylpyrrolidone in 10 days and after control P.
(Boelens et al., 2000) 30 days growth of E. aeruginosa and S.
are used to coat coli, Klebsiella aureus growth rate
silicon shunts for pneumonia was effectively(Giladi et
reduction of S. inhibited (Park et al., al., 2008).
epidermidis. 2003)
• Quorum Quenching
 Quorum quenching is a promising or novel strategy to shut down the virulence expression
(Fig. 4).
 In gram-positive bacteria, the QS inhibitor RNAIII inhibiting peptide (RIP) and halogenated
furanone proved to be very efficacious in preventing and treating staphylococcal infections
associated with CVCs, orthopaedic implants and ureteral stunts (Francolini et al ., 2010).

Fig.4 Prevention of biofilm formation using agents that interfere with QS

(Van der Mei et al., 1988)
 Another drug named as niclosamide reduces the surface motility, biofilm formation and

production of the secreted virulence factors elastase, pyocyanin, and rhamnolipids

(Imperi et al., 2013).

 One more compound cis-9-octadecenoic acid, secreted by Stenotrophomonas

maltophilia which reduces violacein production and biofilm formation by P. aeruginosa

(Singh et al., 2013).

 Various plants, algae, and fungi produce molecules that play a role in inhibiting quorum

sensing in bacteria. For example horseradish-iberin , garlic-ajoene, turmeric-curcumin,

grapefruit extract-furocoumarins, carotenoids, limonoids, pectin, and coumarin are

known to inhibit the QS in bacteria ( Basavaraju et al., 2016).

• Disaggregation of the biofilm matrix
 Different chemicals are used to disaggregate the biofilm ( Fig. 5)

Fig.5 Disaggregation of the biofilm matrix using an enzyme (Van der Mei et al., 1988)
 Dispersin B, a soluble β-N-acetylglucosaminidase purified from A. actinomycetemcomitans
used to prevent staphylococcal biofilm formation (Donelli et al., 2007).
 A treatment with dispersin B followed by a protease (proteinase K or trypsin) can also be able
to destroy biofilms of a Staphylococcal strains on inert surfaces (Chaignon et al., 2007).
 Engineered bacteriophages also able to express Dispersin B and work against E. coli biofilms
(Lu & Collins, 2007) as antibiofilm weapons. They can dissolve the biofilm matrix as well as
kill microbial cells within the biofilm. The use of bacteriophages is a promising approach in
the control of S. epidermidis and P. aeruginosa biofilm formation on catheters (Fu et al.,
2010).
Summary
• Biofilm is an assembly of microorganisms attached to a living surface by a self-
secreted polymeric matrix.
• It is composed of water, microbial cells, either DNA or RNA, polysaccharides and
proteins.
• Biofilm formation is a multi- step process. It includes Attachment, Maturation and
Disaggregation.
• It helps in cleaning the oil spills and treating the waste water.
• It is responsible for causing various diseases like cystic fibrosis, Otitis media,
Periodontitis and other infections.
• These disease effects on death rate of humans. So control and preventive measures
are required.
• Possible antibiofilm strategies can be based on inhibition of sessile growth,
interference with signal molecules and disaggreagtion of biofilms.
References
• Boelens J.J, Tan W.F, Dankert J, Zaat S.A. (2000) Antibacterial activity of antibiotic-
soaked polyvinylpyrrolidone-grafted silicon elastomer hydrocephalus shunts. J Antimicrob Chemoth
45: 221–224.
• Cassinelli C, Morra M, Pavesio A,Renier D.(2000) Evaluation of interfacial properties of hyaluronan
coated poly(methylmethacrylate) intraocular lenses . J Biomat Sci-Polym E 11: 961 – 977.
• Park J.H,Cho Y.W,Choi J.M,ShinH.J, Bae Y.H,Chung H,Jeong S.Y,Know I.C.(2003) Norfloxacin-
releasing urethral catheters for long-term catheterization. J Biomat Sci-Polym E 14: 951–962.
• Van der Mei H.C., Leonard A.J., Weerkamp A.H., Rouxhet P.G & Busscher HJ (1988) .Surface properties
of Streptococcus salivarius HB and nonfibrillar mutants: measurement of zeta potential and elemental
composition with X-ray photoelectron spectroscopy. J Bacteriol; 170: 2462–2466.
• Veerachamy S., Yarlagadda T., Manivasagam G., Yarlagadda P.K (2014). Bacterial adherence and biofilm
formation on medical implants: a review. Proc. Inst. Mech. Eng. H; 228(10):1083–1099.
• Kokare C.R., Kadam S.S.,Mahadik K.R., Chopade B.R., ( 2007) . Studies on bioemulsifier production
from marine Streptomyces spp.SI;Indian J Biotechnol,6 : 78- 84.
• Rabin N., Zheng Y., Opoku-Temeng C., Du Y., Bonsu E., Sintim H O ( 2015).Future Med. Chem. 7(4),
493–51
• Lu T.K,Collins J.J.(2007) Dispersing biofilms with engineered enzymatic bacteriophage. P Natl Acad
Sci USA 104: 11197–11202.
• Imperi F., Massai F., RamachandranP C., Longo F., Zennaro E., Rampioni G., Visca P., Leoni L(2013)
.New life for an old drug: the anthelmintic drug niclosamide inhibits Pseudomonas
aeruginosa quorum sensing.Antimicrob. AgentsChemother. 57:996–1005.
• Jamal M., Tasneem U., Hussain T Andleeb S(2015). Bacterial Biofilm: Its Composition, Formation
and Role in Human Infections. Research & reviews Journal of Microbiology and Biotechnology 4, 3
:1-14.
• Fu W, Forster T, Mayer O, Curtin JJ, Lehman SM & Donlan RM (2010) Bacteriophage cocktail for the
prevention of biofilm formation by Pseudomonas aeruginosa on catheters in an in vitro model
system. Antimicrob Agents Ch 54: 397–404
• Giladi M., Porat Y., Blatt A., Wasserman Y., Kirson E.D., Dekel E., Palti Y(2008). Microbial growth
inhibition by alternating electric fields. Antimicrob. Agents Chemothe ;52(10):3517–3522.
• Francolini I and Donelli G (2010). Prevention and control of biofilm-based medical-device-related
infections.Medical microbiology ;59: 227- 238.
• Basavaraju M.,Sisnity V.V., Palaparthy R., Addanki P. K , Quorum quenching: Signal jamming in
dental plaque biofilms. Journal of Dental Sciences 11, 4: 349-352.
• Biello D, 2015.A review article “ How Microbes Helped Clean BP's Oil Spill”: American scientific